The CRISPR-Cas9 RNA-guided DNA endonuclease has contributed to an explosion of advances in the life sciences that have grown from the ability to edit genomes within living cells.In this Review,we summarize CRISPR-base...The CRISPR-Cas9 RNA-guided DNA endonuclease has contributed to an explosion of advances in the life sciences that have grown from the ability to edit genomes within living cells.In this Review,we summarize CRISPR-based technologies that enable mammalian genome editing and their various applications.展开更多
The rapid development of genome editing technology has brought major breakthroughs in the fields of life science and medicine. In recent years, the clustered regularly interspaced short palindromic repeats(CRISPR)-bas...The rapid development of genome editing technology has brought major breakthroughs in the fields of life science and medicine. In recent years, the clustered regularly interspaced short palindromic repeats(CRISPR)-based genome editing toolbox has been greatly expanded, not only with emerging CRISPR-associated protein(Cas) nucleases, but also novel applications through combination with diverse effectors. Recently, transposon-associated programmable RNA-guided genome editing systems have been uncovered, adding myriads of potential new tools to the genome editing toolbox. CRISPR-based genome editing technology has also revolutionized cardiovascular research. Here we first summarize the advances involving newly identified Cas orthologs, engineered variants and novel genome editing systems, and then discuss the applications of the CRISPR-Cas systems in precise genome editing, such as base editing and prime editing. We also highlight recent progress in cardiovascular research using CRISPR-based genome editing technologies, including the generation of genetically modified in vitro and animal models of cardiovascular diseases(CVD) as well as the applications in treating different types of CVD. Finally, the current limitations and future prospects of genome editing technologies are discussed.展开更多
Genome editing is a valuable tool to target specific DNA sequences for mutagenesis in the genomes of microbes, plants, and animals. Although different genome editing technologies are available, the clustered regularly...Genome editing is a valuable tool to target specific DNA sequences for mutagenesis in the genomes of microbes, plants, and animals. Although different genome editing technologies are available, the clustered regularly interspaced short palindromic repeats/Cas9 （CRISPR/ Cas9） system, which utilizes engineered endonucleases to generate a double-stranded DNA break （DSB） in the target DNA region and subsequently stimulates site-specific mutagenesis through DNA repair machineries, is emerging as a powerful genome editing tool for elucidating mecha- nisms of protection from plant viruses, plant disease resistance, and gene functions in basic and applied research. In this review, we provide an overview of recent advances in the CRISPR system associated genome editing in plants by focusing on application of this technology in model plants, crop plants, fruit plants, woody plants and grasses and discuss how genome editing associated with the CRISPR system can provide insights into genome modifications and functional genomics in plants.展开更多
Targeted mutagenesis based on homologous recombination has been a powerful tool for understanding the mechanisms underlying development, normal physiology, and disease. A recent breakthrough in genome engineering tech...Targeted mutagenesis based on homologous recombination has been a powerful tool for understanding the mechanisms underlying development, normal physiology, and disease. A recent breakthrough in genome engineering technology based on the class of RNA-guided endonucleases, such as clustered regularly interspaced short palindromic repeats(CRISPR)-associated Cas9, is further revolutionizing biology and medical studies. The simplicity of the CRISPR-Cas9 system has enabled its widespread applications in generating germline animal models, somatic genome engineering, and functional genomic screening and in treating genetic and infectious diseases. This technology will likely be used in all fields of biomedicine, ranging from basic research to human gene therapy.展开更多
Human Myoblast Genome Therapy (HMGT) is a platform technology of cell transplantation, nuclear transfer, and tissue engineering. Unlike stem cells, myoblasts are differentiated, immature cells destined to become muscl...Human Myoblast Genome Therapy (HMGT) is a platform technology of cell transplantation, nuclear transfer, and tissue engineering. Unlike stem cells, myoblasts are differentiated, immature cells destined to become muscles. Myoblasts cultured from satellite cells of adult muscle biopsies survive, develop, and function to revitalize degenerative muscles upon transplantation. Injection injury activates regeneration of host myofibers that fuse with the engrafted myoblasts, sharing their nuclei in a common gene pool of the syncytium. Thus, through nuclear transfer and complementation, the normal human genome can be transferred into muscles of patients with genetic disorders to achieve phenotype repair or disease prevention. Myoblasts are safe and efficient gene transfer vehicles endogenous to muscles that constitute 50% of body weight. Results of over 280 HMGT procedures on Duchenne Muscular Dystrophy (DMD) subjects in the past 15 years demonstrated absolute safety. Myoblast-injected DMD muscles showed improved histology. Strength increase at 18 months post-operatively averaged 123%. In another application of HMGT on ischemic cardiomyopathy, the first human myoblast transfer into porcine myocardium revealed that it was safe and effective. Clinical trials on approximately 220 severe cardiomyopathy patients in 15 countries showed a <10% mortality. Most subjects received autologous cells implanted on the epicardial surface during coronory artery bypass graft, or injected on the endomyocardial surface percutaneously through guiding catheters. Significant increases in left ventricular ejection fraction, wall thickness, and wall motion have been reported, with reduction in perfusion defective areas, angina, and shortness of breath. As a new modality of treatment for disease in the skeletal muscle or myocardium, HMGT emerged as safe and effective. Large randomized multi-center trials are under way to confirm these preliminary results. The future of HMGT is bright and exciting.展开更多
The genus Hippophae includes deciduous shrubs or small trees,which provide many ecological,economic,and social benefi ts.We assembled and annotated the chloroplast genomes of sympatric Hippophae gyantsensis(Rousi)Lian...The genus Hippophae includes deciduous shrubs or small trees,which provide many ecological,economic,and social benefi ts.We assembled and annotated the chloroplast genomes of sympatric Hippophae gyantsensis(Rousi)Lian and Hippophae rhamnoides Linn subsp.yunnanensis Rousi and comparatively analyzed their sequences.The fulllength chloroplast genomes of H.gyantsensis and H.rhamnoides subsp.yunnanensis were 155,260 and 156,415 bp,respectively;both featured a quadripartite structure with two copies of a large inverted repeat(IR)separated by small(SSC)and large(LSC)single-copy regions.Each Hippophae chloroplast genome contained 131 genes,comprising 85 protein-coding,8 ribosomal RNA,and 38 transfer RNA genes.Of 1302 nucleotide substitutions found between these twogenomes,824(63.29%)occurred in the intergenic region or intron sequences,and 478(36.71%)were located in the coding sequences.The SSC region had the highest mutation rate,followed by the LSC region and IR regions.Among the protein-coding genes,three had a ratio of nonsynonymous to synonymous substitutions(Ka/Ks)>1 yet none were signifi-cant,and 66 had Ka/Ks<1,of which 46 were signifi cant.We found 20 and 16 optimal codons,most of which ended with A or U,for chloroplast protein-coding genes of H.gyantsensis and H.rhamnoides subsp.yunnanensis,respectively.Phylogenetic analysis of fi ve available whole chloroplast genome sequences in the family Elaeagnaceae—using one Ziziphus jujube sequence as the outgroup—revealed that all fi ve plant species formed a monophyletic clade with two subclades:one subclade consisted of three Hippophae species,while the other was formed by two Elaeagnus species,supported by 100%bootstrap values.Together,these results suggest the chloroplast genomes among Hippophae species are conserved,both in structure and gene composition,due to general purifying selection;like many other plants,a signifi cant AT preference was discerned for most proteincoding genes in the Hippophae chloroplast genome.This study provides a valuable reference tool for future research on the general characteristics and evolution of chloroplast genomes in the genus Hippophae.展开更多
Ginkgo biloba is a famous living“fossil”and has played an important role in the evolution of the Plant Kingdom.Here,the complete chloroplast genome of G.biloba was sequenced and analysed.The chloroplast genome was 1...Ginkgo biloba is a famous living“fossil”and has played an important role in the evolution of the Plant Kingdom.Here,the complete chloroplast genome of G.biloba was sequenced and analysed.The chloroplast genome was 156,990 bp long and predicted to encode 134 genes including 85 protein-coding genes,41 tRNA genes and 8 rRNA genes.The chloroplast genome has a typical quadripartite structure with a pair of inverted repeat regions(IRa and IRb,17,732 bp),a large(LSC,99,259 bp)and small single(SSC,22,267 bp)copy region.After an extensive comparison to previously published gymnosperm plastomes,the gene content and organisation of G.biloba showed high divergence,although part was relatively conserved.The two typical IR regions in the G.biloba chloroplast genome were relatively shorter because it the ycf2 gene.In addition,it was obvious that the IR regions and gene loss were responsible for changes in chloroplast genome size and structure stability,which influenced plastome evolution in different gymnosperms.Phylogenetic analysis revealed that G.biloba is sister to cycads rather than to gnetophytes,cupressophytes,and Pinaceae.Overall,the study showed that the genomic characteristics of G.biloba would be of great help in the further research on the taxonomy,species identification and evolutionary history of gymnosperms,especially for their position in plant systematics and evolution.展开更多
Populus alba × P.glandulosa clone 84 K,derived from South Korea,is widely cultivated in China and used as a model in the molecular research of woody plants because of hi gh gene transformation efficiency.Here,we ...Populus alba × P.glandulosa clone 84 K,derived from South Korea,is widely cultivated in China and used as a model in the molecular research of woody plants because of hi gh gene transformation efficiency.Here,we combined63-fold coverage Illumina short reads and 126-fold coverage PacBio long reads to assemble the genome.Due to the hi gh heterozygosity level at 2.1% estimated by k-mer analysis,we exploited TrioCanu for genome assembly.The PacBio clean subreads of P.alba × P.glandulosa were separated into two parts according to the similarities,compared with the parental genomes of P.alba and P.glandulosa.The two parts of the subreads were assembled to two sets of subgenomes comprising subgenome A(405.31 Mb,from P.alba)and subgenome G(376.05 Mb,from P.glandulosa) with the contig N50 size of 5.43 Mb and 2.15 Mb,respectively.A high-quality P.alba × P.glandulosa genome assembly was obtained.The genome size was 781.36 Mb with the contig N50 size of 3.66 Mb and the longest contig was 19.47 Mb.In addition,a total of 176.95 Mb(43.7%),152.37 Mb(40.5%)of repetitive elements were identified and a total of 38,701 and 38,449 protein-coding genes were predicted in subgenomes A and G,respectively.For functional annotation,96.98% of subgenome A and 96.96% of subgenome G genes were annotated with public databases.This de novo assembled genome will facilitate systematic and comprehensive study,such as multi-omics analysis,in the model tree P.alba X P.glandulosa.展开更多
Quercus L.has significant societal,ecological and economic benefits in the Northern Hemisphere.However,species identification among oaks is notoriously difficult.China harbours highly diverse oaks,of which the diversi...Quercus L.has significant societal,ecological and economic benefits in the Northern Hemisphere.However,species identification among oaks is notoriously difficult.China harbours highly diverse oaks,of which the diversity of white oaks is the most extensive;however,to date,the evolution of chloroplast(cp)genomes in white oaks in China has not been comprehensively studied.Thus,we sequenced the complete cp genomes(161,254 bp,161,229 bp and 161,254 bp in size)of three white oak species(Quercus serrata Thunb.var.brevipetiolata A.DC.Nakai,Quercus wutaishansea Mary and Quercus mongolica Fischer ex Ledebour,respectively).Six white oak species(Quercus aliena Blume,Quercus dentata Thunb.,Quercus aliena Blume var.acutiserrata Maximowicz ex Wenzig,Q.serrata var.brevipetiolata,Q.wutaishanica and Q.mongolica)and five other Fagaceae species(Quercus rubra L.,Quercus variabilis Bl.,Quercus aquifolioid.es Rehd.et Wils.,Fagus engleriana Seem.and Castanea henryi Skan Rehd.et Wils.)were retrieved for comparative analyses.We detected11 highly divergent regions(psbA,matK/rps16,rps16,trnSGCU/trnG-GCC,trnR-UCU/atpA,trnT-GGU/psbD,ndhJ,ndhJ/ndhK,accD,ndhF and ycfl)through comparative analyses and these regions might be used as molecular markers.Theωratio of the rps12,rpoC2 and ycf1 genes was greater than 1 in several comparison groups between white oaks and the petA gene was subjected to significant positive selection between the comparison of six white oaks and Q.variabilis.Phylogenetic analyses revealed that six white oaks were grouped with Q.rubra,forming a single clade.展开更多
Five Larix species(L.griffithii,L.speciose,L.himalaica,L.kongboensis,and L.potaninii var.australis),have survived on the Qinghai-Tibet Plateau(QTP)under specific climate conditions for decades.The lack of genomic info...Five Larix species(L.griffithii,L.speciose,L.himalaica,L.kongboensis,and L.potaninii var.australis),have survived on the Qinghai-Tibet Plateau(QTP)under specific climate conditions for decades.The lack of genomic information seriously hinders research on the evolution,conservation and ecology of these Larix resources.In this study,complete chloroplast(cp)genomes of the 5 species were assembled and compared based on next generation sequencing technology combined with polymerase chain reaction validation.The results show that the 5 cp genomes are relatively conservative in size,gene content and arrangement,and border variation.Phylogenetic analysis showed that the species are closely related as well as to seven other species of the same genus.In addition,the 5 cp genomes contained few simple sequence repeats and relatively low nucleotide variability;thus,12 candidate polymorphic cp DNA markers will be helpful for further research on relevant population genetics.These results will provide valuable genetic information for the conservation,evolution and ecology of these species and their relatives.展开更多
Agrobacterium tumefaciens strain C58 is a type of pathogen that can cause tumors in some dicotyledonous plants.Ever since the genome of A. tumefaciens strain C58 was sequenced, the quality of annotation of its protein...Agrobacterium tumefaciens strain C58 is a type of pathogen that can cause tumors in some dicotyledonous plants.Ever since the genome of A. tumefaciens strain C58 was sequenced, the quality of annotation of its protein-coding genes has been queried continually, because the annotation varies greatly among different databases. In this paper, the questionable hypothetical genes were re-predicted by integrating the TN curve and Z curve methods. As a result, 30 genes originally annotated as ＂hypothetical＂ were discriminated as being non-coding sequences. By testing the re-prediction program 10 times on data sets composed of the function-known genes, the mean accuracy of 99.99% and mean Matthews correlation coefficient value of 0.9999 were obtained. Further sequence analysis and COG analysis showed that the re-annotation results were very reliable. This work can provide an efficient tool and data resources for future studies of A. tumefaciens strain C58.展开更多
The microbial potential of Penicillium has received critical attention.The present research aimed to elucidate the efficacy of crude enzyme secreted from Penicillium oxalicum WX-209 in degrading citrus segments and ev...The microbial potential of Penicillium has received critical attention.The present research aimed to elucidate the efficacy of crude enzyme secreted from Penicillium oxalicum WX-209 in degrading citrus segments and evaluate the safety of the process.Results showed that citrus segment membranes gradually dissolved after treatment with the crude enzyme solution,indicating good degradation capability.No significant differences in body weight,food ingestion rate,hematology,blood biochemistry,and weight changes of different organs were found between the enzyme intake and control groups.Serial experiments showed that the crude enzyme had high biological safety.Moreover,the whole genome of P.oxalicum WX-209 was sequenced by PacBio and Illumina platforms.Twenty-five scaffolds were assembled to generate 36 Mbp size of genome sequence comprising 11369 predicted genes modeled with a GC content of 48.33%.A total of 592 genes were annotated to encode enzymes related to carbohydrates,and some degradation enzyme genes were identified in strain P.oxalicum WX-209.展开更多
Genomics research of Populus deltoides,an important timber species that is widely planted worldwide,is an important part of poplar breeding.Currently,the nuclear and chloroplast genome of P.deltoides have been sequenc...Genomics research of Populus deltoides,an important timber species that is widely planted worldwide,is an important part of poplar breeding.Currently,the nuclear and chloroplast genome of P.deltoides have been sequenced,but its mitochondrial genome(mitogenome)has not been reported.To further explore the evolution and phylogeny of P.deltoides,the mitogenome of P.deltoides I-69 was assembled using reads from Nanopore and Illumina sequencing platforms and found to consist of 802,637 bp and three circular chromosomes(336,205,280,841,and 185,591 bp)containing 58 genes(34 protein-coding genes,21 tRNA genes,and 3 rRNA genes).RNA analysis in combination with several species showed signifi cantly fewer RNA editingsites in the mitogenomes of poplar and other angiosperms than in gymnosperms.Sequence transfer analysis showed extensive mitogenome rearrangements in Populus species,and with evolution from lower to higher plants,tRNA transfer from chloroplasts to mitochondria became increasingly frequent.In a phylogenetic analysis,the evolutionary status of P.deltoides was determined,and the section Populus was supported.Our results based on the fi rst report of a multicircular conformation of the Populus mitogenome provide a basis for further study of the evolution and genetics of P.deltoides and other Populus species and for breeding programs.展开更多
The effect of Ar^＋ beam implantation and maize genome DNA on autotetraploid rice is studied. Better mutation types and higher mutation rates were discovered in M2 of T3 with ion implantation and immersion in maize ge...The effect of Ar^＋ beam implantation and maize genome DNA on autotetraploid rice is studied. Better mutation types and higher mutation rates were discovered in M2 of T3 with ion implantation and immersion in maize genome DNA. In the five agronomic categories investigated, the mutation rate of the seed setting rate was 9.1%, and the total mutation rate was 14.8% in the T3. However, the total mutation rate was 2.1% with the treatment of only ion implantation and 1.3% with the treatment of only immersion in maize genome DNA. Mutant FA36（4） was discovered in M1 with ion beam implantation and immersion in maize genome DNA. Its RuBPCase activity, PEPCase activity and seed setting rate were 32%, 153%, and 36.79%, respectively, higher than its parent IR36（4）. Rapid analysis of polymorphicDNA （RAPD） analysis of three M2 plants of FA36（4） （FMI, FM2, FM3） and two controls （purple maize and IR36（4）） were also conducted with 40 random primers. S5-3 was RAPD fragment amplified with a template of purple maize, FM2 and FM3 genome DNA using primer S5. There was no S5-3 in the RAPD pattern of IR36（4） or FMI.展开更多
High-throughput computational materials design provides one efficient solution to accelerate the discovery and development of functional materials. Its core concept is to build a large quantum materials repository and...High-throughput computational materials design provides one efficient solution to accelerate the discovery and development of functional materials. Its core concept is to build a large quantum materials repository and to search for target materials with desired properties via appropriate materials descriptors in a high-throughput fashion, which shares the same idea with the materials genome approach. This article reviews recent progress of discovering and developing new functional materials using high-throughput computational materials design approach. Emphasis is placed on the rational design of high-throughput screening procedure and the development of appropriate materials descriptors, concentrating on the electronic and magnetic properties of functional materials for various types of industrial applications in nanoelectronics.展开更多
Molecular genetic maps were commonly constructed by analyzing the segregation of restriction fragment length polymorphisms (RFLPs). Here we described methodology-marker sequences in a new mapping based on recent docum...Molecular genetic maps were commonly constructed by analyzing the segregation of restriction fragment length polymorphisms (RFLPs). Here we described methodology-marker sequences in a new mapping based on recent documents. With the methods they were unique sequences detected by the polymerase chain reaction (PCR). Each of the methods had its Iimitations and the current trend was to integrate the maps produced by the different methods. Marker sequences contained mainly expressed sequence tags (ESTs),polymorphie sequence-tagged sites (STSs), randomly amplified polymorphic DNA (RAPDs), cIeaved amplified polymorphic sequences (CAPS), amplified fragment Iength pofymorphism (AFLPs), genorne sequence sampling (GSS) and sequence-tagged connectors (STCs) in this paper.展开更多
Species of the Pinus genus provide a classical model for studying hybrid speciation.Although studies on two narrowly distributed species(P inus funebris and P.takahasii)concluded that they originated from two widespre...Species of the Pinus genus provide a classical model for studying hybrid speciation.Although studies on two narrowly distributed species(P inus funebris and P.takahasii)concluded that they originated from two widespread species(P.sylvestris and P.densiflora)via hybrid speciation,the conclusion was based on a low number of informative restriction sites.In this study,we analyzed the sequences of four Pinus chloroplast(cp)genomes(P.sylvestris,P.densiflora,P.funebris and P.takahasii)to clarify whether hybrid speciation was involved.The complete cp-genomes of Pinus species ranged in size from 119,865 to 119,890 bp,similar to other Pinus species.Phylogenetic results based on the whole cp-genomes showed P.sylvestris clustered with P.funebris and P.takahasii,which suggested that P.sylvestris was the paternal parent in hybridization events.In an analysis of simple sequence repeats(SSRs),we detected a total of 69 SSRs repeats among the four Pinus cp-genomes;most were A or T bases.In addition,we identified divergent hotspot regions among the four Pinus cp-genomes(trnE-clpP,cemA-ycf4,petD-rpoA,psbD-trnT,and trnN-chlL),in P.sylvestris(psbD-trnT,trnN-chlL,psbB and rps8)and in P.densiflora(trnE-clpP,petD-rpoA,ycf3 intron,psbD-trnT,and trnN-chlL).The genome information found in this study provides new insights into hybrid speciation in P inus and contributes to a better understanding of the phylogenetic relationships within the Pinus genus.展开更多
objective: To study the effects of human genome DNA on the cultured spinal cord neurons of em bryonic mouse. Methods: The human genome DNA was added to the culture medium of the spinal cord neu rons of embryonic mouse...objective: To study the effects of human genome DNA on the cultured spinal cord neurons of em bryonic mouse. Methods: The human genome DNA was added to the culture medium of the spinal cord neu rons of embryonic mouse. Eight days later, MTT assay, NSE immunocytochemical staining and image analy sis were proformed to examine the viabilities and the neurites lengths of the neurons. Results: The neurite length of the experimental group was significantly Ionger than that of the control group, but no marked dif ference was found between the viabilities of the neurons of the experimental groups and that of the control ones. Conclusiou: Human genome DNA has no effects on the viabilities of the cultured neurons but can pro mote the neurite growth.展开更多
文摘The CRISPR-Cas9 RNA-guided DNA endonuclease has contributed to an explosion of advances in the life sciences that have grown from the ability to edit genomes within living cells.In this Review,we summarize CRISPR-based technologies that enable mammalian genome editing and their various applications.
基金supported by the National Natural Science Foundation of China (82270355, 82270354, 81970134, 82030011, 31630093)the National Key Research and Development Program of China (2019YFA0801601, 2021YFA1101801)。
文摘The rapid development of genome editing technology has brought major breakthroughs in the fields of life science and medicine. In recent years, the clustered regularly interspaced short palindromic repeats(CRISPR)-based genome editing toolbox has been greatly expanded, not only with emerging CRISPR-associated protein(Cas) nucleases, but also novel applications through combination with diverse effectors. Recently, transposon-associated programmable RNA-guided genome editing systems have been uncovered, adding myriads of potential new tools to the genome editing toolbox. CRISPR-based genome editing technology has also revolutionized cardiovascular research. Here we first summarize the advances involving newly identified Cas orthologs, engineered variants and novel genome editing systems, and then discuss the applications of the CRISPR-Cas systems in precise genome editing, such as base editing and prime editing. We also highlight recent progress in cardiovascular research using CRISPR-based genome editing technologies, including the generation of genetically modified in vitro and animal models of cardiovascular diseases(CVD) as well as the applications in treating different types of CVD. Finally, the current limitations and future prospects of genome editing technologies are discussed.
文摘Genome editing is a valuable tool to target specific DNA sequences for mutagenesis in the genomes of microbes, plants, and animals. Although different genome editing technologies are available, the clustered regularly interspaced short palindromic repeats/Cas9 （CRISPR/ Cas9） system, which utilizes engineered endonucleases to generate a double-stranded DNA break （DSB） in the target DNA region and subsequently stimulates site-specific mutagenesis through DNA repair machineries, is emerging as a powerful genome editing tool for elucidating mecha- nisms of protection from plant viruses, plant disease resistance, and gene functions in basic and applied research. In this review, we provide an overview of recent advances in the CRISPR system associated genome editing in plants by focusing on application of this technology in model plants, crop plants, fruit plants, woody plants and grasses and discuss how genome editing associated with the CRISPR system can provide insights into genome modifications and functional genomics in plants.
基金supported by the Chinese National Key Program on Basic Research (2012CB945103, 2011CB504202)National Natural Science Foundation of China (31430057)
文摘Targeted mutagenesis based on homologous recombination has been a powerful tool for understanding the mechanisms underlying development, normal physiology, and disease. A recent breakthrough in genome engineering technology based on the class of RNA-guided endonucleases, such as clustered regularly interspaced short palindromic repeats(CRISPR)-associated Cas9, is further revolutionizing biology and medical studies. The simplicity of the CRISPR-Cas9 system has enabled its widespread applications in generating germline animal models, somatic genome engineering, and functional genomic screening and in treating genetic and infectious diseases. This technology will likely be used in all fields of biomedicine, ranging from basic research to human gene therapy.
文摘Human Myoblast Genome Therapy (HMGT) is a platform technology of cell transplantation, nuclear transfer, and tissue engineering. Unlike stem cells, myoblasts are differentiated, immature cells destined to become muscles. Myoblasts cultured from satellite cells of adult muscle biopsies survive, develop, and function to revitalize degenerative muscles upon transplantation. Injection injury activates regeneration of host myofibers that fuse with the engrafted myoblasts, sharing their nuclei in a common gene pool of the syncytium. Thus, through nuclear transfer and complementation, the normal human genome can be transferred into muscles of patients with genetic disorders to achieve phenotype repair or disease prevention. Myoblasts are safe and efficient gene transfer vehicles endogenous to muscles that constitute 50% of body weight. Results of over 280 HMGT procedures on Duchenne Muscular Dystrophy (DMD) subjects in the past 15 years demonstrated absolute safety. Myoblast-injected DMD muscles showed improved histology. Strength increase at 18 months post-operatively averaged 123%. In another application of HMGT on ischemic cardiomyopathy, the first human myoblast transfer into porcine myocardium revealed that it was safe and effective. Clinical trials on approximately 220 severe cardiomyopathy patients in 15 countries showed a <10% mortality. Most subjects received autologous cells implanted on the epicardial surface during coronory artery bypass graft, or injected on the endomyocardial surface percutaneously through guiding catheters. Significant increases in left ventricular ejection fraction, wall thickness, and wall motion have been reported, with reduction in perfusion defective areas, angina, and shortness of breath. As a new modality of treatment for disease in the skeletal muscle or myocardium, HMGT emerged as safe and effective. Large randomized multi-center trials are under way to confirm these preliminary results. The future of HMGT is bright and exciting.
基金the National Natural Science Foundation of China(31670666)the Fundamental Research Funds for the Central Non-profit Research Institution of Chinese Academy of Forestry(ZDRIF201706).
文摘The genus Hippophae includes deciduous shrubs or small trees,which provide many ecological,economic,and social benefi ts.We assembled and annotated the chloroplast genomes of sympatric Hippophae gyantsensis(Rousi)Lian and Hippophae rhamnoides Linn subsp.yunnanensis Rousi and comparatively analyzed their sequences.The fulllength chloroplast genomes of H.gyantsensis and H.rhamnoides subsp.yunnanensis were 155,260 and 156,415 bp,respectively;both featured a quadripartite structure with two copies of a large inverted repeat(IR)separated by small(SSC)and large(LSC)single-copy regions.Each Hippophae chloroplast genome contained 131 genes,comprising 85 protein-coding,8 ribosomal RNA,and 38 transfer RNA genes.Of 1302 nucleotide substitutions found between these twogenomes,824(63.29%)occurred in the intergenic region or intron sequences,and 478(36.71%)were located in the coding sequences.The SSC region had the highest mutation rate,followed by the LSC region and IR regions.Among the protein-coding genes,three had a ratio of nonsynonymous to synonymous substitutions(Ka/Ks)>1 yet none were signifi-cant,and 66 had Ka/Ks<1,of which 46 were signifi cant.We found 20 and 16 optimal codons,most of which ended with A or U,for chloroplast protein-coding genes of H.gyantsensis and H.rhamnoides subsp.yunnanensis,respectively.Phylogenetic analysis of fi ve available whole chloroplast genome sequences in the family Elaeagnaceae—using one Ziziphus jujube sequence as the outgroup—revealed that all fi ve plant species formed a monophyletic clade with two subclades:one subclade consisted of three Hippophae species,while the other was formed by two Elaeagnus species,supported by 100%bootstrap values.Together,these results suggest the chloroplast genomes among Hippophae species are conserved,both in structure and gene composition,due to general purifying selection;like many other plants,a signifi cant AT preference was discerned for most proteincoding genes in the Hippophae chloroplast genome.This study provides a valuable reference tool for future research on the general characteristics and evolution of chloroplast genomes in the genus Hippophae.
基金supported by the Key Forestry Public Welfare Project of China(201504105)the National Key Research and Development Program of China(2017YFD0600700)the Agricultural Science and Technology Independent Innovation Funds of Jiangsu Province[CX(16)1005].
文摘Ginkgo biloba is a famous living“fossil”and has played an important role in the evolution of the Plant Kingdom.Here,the complete chloroplast genome of G.biloba was sequenced and analysed.The chloroplast genome was 156,990 bp long and predicted to encode 134 genes including 85 protein-coding genes,41 tRNA genes and 8 rRNA genes.The chloroplast genome has a typical quadripartite structure with a pair of inverted repeat regions(IRa and IRb,17,732 bp),a large(LSC,99,259 bp)and small single(SSC,22,267 bp)copy region.After an extensive comparison to previously published gymnosperm plastomes,the gene content and organisation of G.biloba showed high divergence,although part was relatively conserved.The two typical IR regions in the G.biloba chloroplast genome were relatively shorter because it the ycf2 gene.In addition,it was obvious that the IR regions and gene loss were responsible for changes in chloroplast genome size and structure stability,which influenced plastome evolution in different gymnosperms.Phylogenetic analysis revealed that G.biloba is sister to cycads rather than to gnetophytes,cupressophytes,and Pinaceae.Overall,the study showed that the genomic characteristics of G.biloba would be of great help in the further research on the taxonomy,species identification and evolutionary history of gymnosperms,especially for their position in plant systematics and evolution.
基金supported by grants CAFYBB2017ZY001 and TGB2016001 from Fundamental Research Funds of the Chinese Academy of Forestry。
文摘Populus alba × P.glandulosa clone 84 K,derived from South Korea,is widely cultivated in China and used as a model in the molecular research of woody plants because of hi gh gene transformation efficiency.Here,we combined63-fold coverage Illumina short reads and 126-fold coverage PacBio long reads to assemble the genome.Due to the hi gh heterozygosity level at 2.1% estimated by k-mer analysis,we exploited TrioCanu for genome assembly.The PacBio clean subreads of P.alba × P.glandulosa were separated into two parts according to the similarities,compared with the parental genomes of P.alba and P.glandulosa.The two parts of the subreads were assembled to two sets of subgenomes comprising subgenome A(405.31 Mb,from P.alba)and subgenome G(376.05 Mb,from P.glandulosa) with the contig N50 size of 5.43 Mb and 2.15 Mb,respectively.A high-quality P.alba × P.glandulosa genome assembly was obtained.The genome size was 781.36 Mb with the contig N50 size of 3.66 Mb and the longest contig was 19.47 Mb.In addition,a total of 176.95 Mb(43.7%),152.37 Mb(40.5%)of repetitive elements were identified and a total of 38,701 and 38,449 protein-coding genes were predicted in subgenomes A and G,respectively.For functional annotation,96.98% of subgenome A and 96.96% of subgenome G genes were annotated with public databases.This de novo assembled genome will facilitate systematic and comprehensive study,such as multi-omics analysis,in the model tree P.alba X P.glandulosa.
基金the Fundamental Research Funds for the Central Non-Profit Research Institution of CAF(CAFYBB2018ZB001)the National Natural Science Foundation of China(42071065)。
文摘Quercus L.has significant societal,ecological and economic benefits in the Northern Hemisphere.However,species identification among oaks is notoriously difficult.China harbours highly diverse oaks,of which the diversity of white oaks is the most extensive;however,to date,the evolution of chloroplast(cp)genomes in white oaks in China has not been comprehensively studied.Thus,we sequenced the complete cp genomes(161,254 bp,161,229 bp and 161,254 bp in size)of three white oak species(Quercus serrata Thunb.var.brevipetiolata A.DC.Nakai,Quercus wutaishansea Mary and Quercus mongolica Fischer ex Ledebour,respectively).Six white oak species(Quercus aliena Blume,Quercus dentata Thunb.,Quercus aliena Blume var.acutiserrata Maximowicz ex Wenzig,Q.serrata var.brevipetiolata,Q.wutaishanica and Q.mongolica)and five other Fagaceae species(Quercus rubra L.,Quercus variabilis Bl.,Quercus aquifolioid.es Rehd.et Wils.,Fagus engleriana Seem.and Castanea henryi Skan Rehd.et Wils.)were retrieved for comparative analyses.We detected11 highly divergent regions(psbA,matK/rps16,rps16,trnSGCU/trnG-GCC,trnR-UCU/atpA,trnT-GGU/psbD,ndhJ,ndhJ/ndhK,accD,ndhF and ycfl)through comparative analyses and these regions might be used as molecular markers.Theωratio of the rps12,rpoC2 and ycf1 genes was greater than 1 in several comparison groups between white oaks and the petA gene was subjected to significant positive selection between the comparison of six white oaks and Q.variabilis.Phylogenetic analyses revealed that six white oaks were grouped with Q.rubra,forming a single clade.
基金the National Natural Science Foundation of China(31660215)the Construction Project for First-Class Ecology Discipline in Guizhou(GNYL[2017]007),ChinaMajor Scientifi c and Technological Projects of Guizhou Province([2018]5261),China。
文摘Five Larix species(L.griffithii,L.speciose,L.himalaica,L.kongboensis,and L.potaninii var.australis),have survived on the Qinghai-Tibet Plateau(QTP)under specific climate conditions for decades.The lack of genomic information seriously hinders research on the evolution,conservation and ecology of these Larix resources.In this study,complete chloroplast(cp)genomes of the 5 species were assembled and compared based on next generation sequencing technology combined with polymerase chain reaction validation.The results show that the 5 cp genomes are relatively conservative in size,gene content and arrangement,and border variation.Phylogenetic analysis showed that the species are closely related as well as to seven other species of the same genus.In addition,the 5 cp genomes contained few simple sequence repeats and relatively low nucleotide variability;thus,12 candidate polymorphic cp DNA markers will be helpful for further research on relevant population genetics.These results will provide valuable genetic information for the conservation,evolution and ecology of these species and their relatives.
基金Project supported by the National Natural Science Foundation of China(Grant Nos.61302186 and 61271378)the Funding from the State Key Laboratory of Bioelectronics of Southeast University
文摘Agrobacterium tumefaciens strain C58 is a type of pathogen that can cause tumors in some dicotyledonous plants.Ever since the genome of A. tumefaciens strain C58 was sequenced, the quality of annotation of its protein-coding genes has been queried continually, because the annotation varies greatly among different databases. In this paper, the questionable hypothetical genes were re-predicted by integrating the TN curve and Z curve methods. As a result, 30 genes originally annotated as ＂hypothetical＂ were discriminated as being non-coding sequences. By testing the re-prediction program 10 times on data sets composed of the function-known genes, the mean accuracy of 99.99% and mean Matthews correlation coefficient value of 0.9999 were obtained. Further sequence analysis and COG analysis showed that the re-annotation results were very reliable. This work can provide an efficient tool and data resources for future studies of A. tumefaciens strain C58.
基金the financial support of the National Natural Science Foundation of China[32201960,32073020]Science and Technology Innovation Program of Hunan Province[2022RC1150]+2 种基金Changsha Municipal Natural Science Foundation[kq2202332]Hunan innovative province construction project[2019NK2041]Agricultural Science and Technology Innovation Project of Hunan Province[2021CX05].
文摘The microbial potential of Penicillium has received critical attention.The present research aimed to elucidate the efficacy of crude enzyme secreted from Penicillium oxalicum WX-209 in degrading citrus segments and evaluate the safety of the process.Results showed that citrus segment membranes gradually dissolved after treatment with the crude enzyme solution,indicating good degradation capability.No significant differences in body weight,food ingestion rate,hematology,blood biochemistry,and weight changes of different organs were found between the enzyme intake and control groups.Serial experiments showed that the crude enzyme had high biological safety.Moreover,the whole genome of P.oxalicum WX-209 was sequenced by PacBio and Illumina platforms.Twenty-five scaffolds were assembled to generate 36 Mbp size of genome sequence comprising 11369 predicted genes modeled with a GC content of 48.33%.A total of 592 genes were annotated to encode enzymes related to carbohydrates,and some degradation enzyme genes were identified in strain P.oxalicum WX-209.
基金funded by the National Key Research and Development Program of China[Grant Number 2021YFD2201205]the Priority Academic Program Development of Jiangsu Higher Education Institutions(PAPD).
文摘Genomics research of Populus deltoides,an important timber species that is widely planted worldwide,is an important part of poplar breeding.Currently,the nuclear and chloroplast genome of P.deltoides have been sequenced,but its mitochondrial genome(mitogenome)has not been reported.To further explore the evolution and phylogeny of P.deltoides,the mitogenome of P.deltoides I-69 was assembled using reads from Nanopore and Illumina sequencing platforms and found to consist of 802,637 bp and three circular chromosomes(336,205,280,841,and 185,591 bp)containing 58 genes(34 protein-coding genes,21 tRNA genes,and 3 rRNA genes).RNA analysis in combination with several species showed signifi cantly fewer RNA editingsites in the mitogenomes of poplar and other angiosperms than in gymnosperms.Sequence transfer analysis showed extensive mitogenome rearrangements in Populus species,and with evolution from lower to higher plants,tRNA transfer from chloroplasts to mitochondria became increasingly frequent.In a phylogenetic analysis,the evolutionary status of P.deltoides was determined,and the section Populus was supported.Our results based on the fi rst report of a multicircular conformation of the Populus mitogenome provide a basis for further study of the evolution and genetics of P.deltoides and other Populus species and for breeding programs.
基金the National Key Program of China(2001BA302B03)Doctoral Startup Foundation of Huuan Science and Technology University of China(E54141)Educational Department Foundation of Hunan Province of China
文摘The effect of Ar^＋ beam implantation and maize genome DNA on autotetraploid rice is studied. Better mutation types and higher mutation rates were discovered in M2 of T3 with ion implantation and immersion in maize genome DNA. In the five agronomic categories investigated, the mutation rate of the seed setting rate was 9.1%, and the total mutation rate was 14.8% in the T3. However, the total mutation rate was 2.1% with the treatment of only ion implantation and 1.3% with the treatment of only immersion in maize genome DNA. Mutant FA36（4） was discovered in M1 with ion beam implantation and immersion in maize genome DNA. Its RuBPCase activity, PEPCase activity and seed setting rate were 32%, 153%, and 36.79%, respectively, higher than its parent IR36（4）. Rapid analysis of polymorphicDNA （RAPD） analysis of three M2 plants of FA36（4） （FMI, FM2, FM3） and two controls （purple maize and IR36（4）） were also conducted with 40 random primers. S5-3 was RAPD fragment amplified with a template of purple maize, FM2 and FM3 genome DNA using primer S5. There was no S5-3 in the RAPD pattern of IR36（4） or FMI.
基金support by National Science Foundation under award number ACI-1550404American Chemical Society Petroleum Research Fund under the award number 55481-DNI6+1 种基金Global Research Outreach(GRO)Program of Samsung Advanced Institute of Technology under the award number 20164974the Vannevar Bush Faculty Fellowship program sponsored by the Basic Research Office of the Assistant Secretary of Defense for Research and Engineering under the Office of Naval Research grant N00014-16-1-2569
文摘High-throughput computational materials design provides one efficient solution to accelerate the discovery and development of functional materials. Its core concept is to build a large quantum materials repository and to search for target materials with desired properties via appropriate materials descriptors in a high-throughput fashion, which shares the same idea with the materials genome approach. This article reviews recent progress of discovering and developing new functional materials using high-throughput computational materials design approach. Emphasis is placed on the rational design of high-throughput screening procedure and the development of appropriate materials descriptors, concentrating on the electronic and magnetic properties of functional materials for various types of industrial applications in nanoelectronics.
文摘Molecular genetic maps were commonly constructed by analyzing the segregation of restriction fragment length polymorphisms (RFLPs). Here we described methodology-marker sequences in a new mapping based on recent documents. With the methods they were unique sequences detected by the polymerase chain reaction (PCR). Each of the methods had its Iimitations and the current trend was to integrate the maps produced by the different methods. Marker sequences contained mainly expressed sequence tags (ESTs),polymorphie sequence-tagged sites (STSs), randomly amplified polymorphic DNA (RAPDs), cIeaved amplified polymorphic sequences (CAPS), amplified fragment Iength pofymorphism (AFLPs), genorne sequence sampling (GSS) and sequence-tagged connectors (STCs) in this paper.
基金funded by Kunyu Mountain National Nature Reserve Administration。
文摘Species of the Pinus genus provide a classical model for studying hybrid speciation.Although studies on two narrowly distributed species(P inus funebris and P.takahasii)concluded that they originated from two widespread species(P.sylvestris and P.densiflora)via hybrid speciation,the conclusion was based on a low number of informative restriction sites.In this study,we analyzed the sequences of four Pinus chloroplast(cp)genomes(P.sylvestris,P.densiflora,P.funebris and P.takahasii)to clarify whether hybrid speciation was involved.The complete cp-genomes of Pinus species ranged in size from 119,865 to 119,890 bp,similar to other Pinus species.Phylogenetic results based on the whole cp-genomes showed P.sylvestris clustered with P.funebris and P.takahasii,which suggested that P.sylvestris was the paternal parent in hybridization events.In an analysis of simple sequence repeats(SSRs),we detected a total of 69 SSRs repeats among the four Pinus cp-genomes;most were A or T bases.In addition,we identified divergent hotspot regions among the four Pinus cp-genomes(trnE-clpP,cemA-ycf4,petD-rpoA,psbD-trnT,and trnN-chlL),in P.sylvestris(psbD-trnT,trnN-chlL,psbB and rps8)and in P.densiflora(trnE-clpP,petD-rpoA,ycf3 intron,psbD-trnT,and trnN-chlL).The genome information found in this study provides new insights into hybrid speciation in P inus and contributes to a better understanding of the phylogenetic relationships within the Pinus genus.
文摘objective: To study the effects of human genome DNA on the cultured spinal cord neurons of em bryonic mouse. Methods: The human genome DNA was added to the culture medium of the spinal cord neu rons of embryonic mouse. Eight days later, MTT assay, NSE immunocytochemical staining and image analy sis were proformed to examine the viabilities and the neurites lengths of the neurons. Results: The neurite length of the experimental group was significantly Ionger than that of the control group, but no marked dif ference was found between the viabilities of the neurons of the experimental groups and that of the control ones. Conclusiou: Human genome DNA has no effects on the viabilities of the cultured neurons but can pro mote the neurite growth.
