Acta Geologica Sinica(English Edition)is a bimonthly,SCI-indexed academic journal produced by the Geological Society of China.With the latest 2022 Impact Factor of 3.3,it has long been indexed by many international da...Acta Geologica Sinica(English Edition)is a bimonthly,SCI-indexed academic journal produced by the Geological Society of China.With the latest 2022 Impact Factor of 3.3,it has long been indexed by many international databases and websites,such as SCI,CA,BIG,etc.Internationally,the journal cooperates with John Wiley&Sons,Inc.to publish the electronic version;all papers can be downloaded online.Contributors wishing to submit,read,enquire and download can log in via http://mc.manuscriptcentral.com/ags.展开更多
“Journal of Jilin University(Science Edition)”is a comprehensive academic journal in the fields of science sponsored by Jilin University and administrated by the Ministry of Education of the People's Republic of...“Journal of Jilin University(Science Edition)”is a comprehensive academic journal in the fields of science sponsored by Jilin University and administrated by the Ministry of Education of the People's Republic of China.The journal started publication in 1955.The original name at starting publication was“Journal of Natural Science of Northeast People University”,which was changed into“Acta Scientiarum Naturalium Universitatis Jilinensis”in 1958 owing to the name change of the university.展开更多
“Journal of Jilin University(Science Edition)” is a comprehensive academic journal in the fields of science sponsored by Jilin University and administrated by the Ministry of Education of the People's Republic o...“Journal of Jilin University(Science Edition)” is a comprehensive academic journal in the fields of science sponsored by Jilin University and administrated by the Ministry of Education of the People's Republic of China.The journal started publication in 1955.The original name at starting publication was “Journal of Natural Science of Northeast People University”,which was changed into “Acta Scientiarum Naturalium Universitatis Jilinensis” in 1958 owing to the name change of the university.展开更多
“Journal of Jilin University(Science Edition)”is a comprehensive academic journal in the fields of science sponsored by Jilin University and administrated by the Ministry of Education of the People's Republic of...“Journal of Jilin University(Science Edition)”is a comprehensive academic journal in the fields of science sponsored by Jilin University and administrated by the Ministry of Education of the People's Republic of China.The journal started publication in 1955.The original name at starting publication was“Journal of Natural Science of Northeast People University”.展开更多
“Journal of Jilin University(Science Edition)” is a comprehensive academic journal in the fields of science sponsored by Jilin University and administrated by the Ministry of Education of the People’s Republic of C...“Journal of Jilin University(Science Edition)” is a comprehensive academic journal in the fields of science sponsored by Jilin University and administrated by the Ministry of Education of the People’s Republic of China.The journal started publication in 1955.The original name at starting publication was “Journal of Natural Science of Northeast People University”.展开更多
“Journal of Jilin University(Science Edition)” is a comprehensive academic journal in the fields of science sponsored by Jilin University and administrated by the Ministry of Education of the People’s Republic of C...“Journal of Jilin University(Science Edition)” is a comprehensive academic journal in the fields of science sponsored by Jilin University and administrated by the Ministry of Education of the People’s Republic of China.The journal started publication in 1955.The original name at starting publication was “Journal of Natural Science of Northeast People University”,which was changed into “Acta Scientiarum Naturalium Universitatis Jilinensis” in 1958 owing to the name change of the university.展开更多
Jute(Corchorus capsularis L.)is the second most important natural plant fiber source after cotton.However,developing an efficient gene editing system for jute remains a challenge.In this study,the transgenic hairy roo...Jute(Corchorus capsularis L.)is the second most important natural plant fiber source after cotton.However,developing an efficient gene editing system for jute remains a challenge.In this study,the transgenic hairy root system mediated by Agrobacterium rhizogenes strain K599 was developed for Meifeng 4,an elite jute variety widely cultivated in China.The transgenic hairy root system for jute was verified by subcellular localization and bimolecular fluorescence complementation(BiFC)assays.The CHLOROPLASTOS ALTERADOS 1(CcCLA1)gene,which is involved in the development of chloroplasts,was targeted for editing at two sites in Meifeng 4.Based on this hairy root transformation,the gRNA scaffold was placed under the control of cotton ubiquitin GhU6.7 and-GhU6.9 promoters,respectively,to assess the efficiency of gene editing.Results indicated the 50.0%(GhU6.7)and 38.5%(GhU6.9)editing events in the target 2 alleles(gRNA2),but no mutation was detected in the target 1 allele(gRNA1)in transgenic-positive hairy roots.CcCLA1 gene editing at gRNA2 under the control of GhU6.7 in Meifeng 4 was also carried out by Agrobacterium tumefaciens-mediated transformation.Two CcCLA1 mutants were albinic,with a gene editing efficiency of 5.3%.These findings confirm that the CRISPR/Cas9 system,incorporating promoter GhU6.7,can be used as a gene editing tool for jute.展开更多
The development and maturation of the CRISPR/Cas genome editing system provides a valuable tool for plant functional genomics and genetic improvement.Currently available genome-editing tools have a limited number of t...The development and maturation of the CRISPR/Cas genome editing system provides a valuable tool for plant functional genomics and genetic improvement.Currently available genome-editing tools have a limited number of targets,restricting their application in genetic research.In this study,we developed a novel CRISPR/Cas9 plant ultra-multiplex genome editing system consisting of two template vectors,eight donor vectors,four destination vectors,and one primer-design software package.By combining the advantages of Golden Gate cloning to assemble multiple repetitive fragments and Gateway recombination to assemble large fragments and by changing the structure of the amplicons used to assemble sg RNA expression cassettes,the plant ultra-multiplex genome editing system can assemble a single binary vector targeting more than 40 genomic loci.A rice knockout vector containing 49 sg RNA expression cassettes was assembled and a high co-editing efficiency was observed.This plant ultra-multiplex genome editing system advances synthetic biology and plant genetic engineering.展开更多
Aims and Scope Journal of Northeast Agricultural University(English Edition)is a comprehensive academic journal on agricultural sciences sponsored by Northeast Agricultural University and distributed worldwide.It is a...Aims and Scope Journal of Northeast Agricultural University(English Edition)is a comprehensive academic journal on agricultural sciences sponsored by Northeast Agricultural University and distributed worldwide.It is a peer reviewed journal published quarterly and mainly publishes review and research articles that reflect the latest achievements on crop science,horticulture,plant protection,resource and environment,animal science,veterinary medicine,agricultural engineering and technology,agricultural water conservancy,life science,biotechnology and food science.展开更多
Aims and Scope Journal of Northeast Agricultural University(English Edition) is a comprehensive academic journal on agricultural sciences sponsored by Northeast Agricultural University and distributed worldwide. It is...Aims and Scope Journal of Northeast Agricultural University(English Edition) is a comprehensive academic journal on agricultural sciences sponsored by Northeast Agricultural University and distributed worldwide. It is a peer reviewed journal published quarterly and mainly publishes review and research articles that reflect the latest achievements on crop science, horticulture, plant protection, resource and environment, animal science, veterinary medicine, agricultural engineering and technology, agricultural water conservancy, life science, biotechnology and food science.展开更多
Aims and Scope Journal of Northeast Agricultural Univer-sity(English Edition)is a comprehensive academic journal on agricultural sciences sponsored by Northeast Agricultural University and distributed worldwide.It is ...Aims and Scope Journal of Northeast Agricultural Univer-sity(English Edition)is a comprehensive academic journal on agricultural sciences sponsored by Northeast Agricultural University and distributed worldwide.It is a peer reviewed journal published quarterly and mainly publishes review and research articles that reflect the latest achievements on crop science,horticulture,plant protection,resource and environment,animal science,veterinary medicine,agricultural engineering and technology,agricultural water conservancy,life science,biotechnology and food science.展开更多
Objective:To validate the eighth edition of the AJCC staging system in patients with pancreatic adenocarcinoma receiving only stereotactic body radiation therapy and chemotherapy,and to propose modifications to improv...Objective:To validate the eighth edition of the AJCC staging system in patients with pancreatic adenocarcinoma receiving only stereotactic body radiation therapy and chemotherapy,and to propose modifications to improve prognostic accuracy.Methods:Patients with pathologically confirmed pancreatic adenocarcinoma without metastasis who were undergoing only chemoradiotherapy were included and staged according to the seventh and eighth editions of the AJCC staging system.Meanwhile,another group of stage T4 patients from the above enrollment with only portal vein involvement with or without tumor thrombi(PV±PVTT)were retrieved for survival comparisons.Modifications were proposed according to the survival comparisons.A cohort from the SEER database was used for external validation of the modified staging system.Results:A total of 683 patients were included.Patients with N2 or N1 but different T stages had significantly different survival outcomes according to the eighth edition.The survival of patients with(PV±PVTT)was comparable to that of patients with T4 tumors.The concordance index of the seventh and eighth editions,and the modified staging system was 0.744(95%CI:0.718—0.769),0.750(95%CI:0.725—0.775),and 0.788(95%CI:0.762-0.813),respectively.For external validation,the concordance index was 0.744(95%CI:0.718-0.770),0.750(95%CI:0.724-0.776),and 0.788(95%CI:0.762-0.814),respectively.Conclusions:The modified staging system is suggested to have the m ost accurate prognostic value.Hence,PV土PVTT should be added to the definition of T4 tumors regardless of tumor size.Patients with N2 or N1 in different T stages could be regrouped into different substages.Additionally,stage III should be subclassified into IIIA(T3N 2 and T4N 0)and IIIB(T4N 1-2).展开更多
Pea(Pisum sativum L.)is an annual cool-season legume crop.Owing to its role in sustainable agriculture as both a rotation and a cash crop,its global market is expanding and increased production is urgently needed.For ...Pea(Pisum sativum L.)is an annual cool-season legume crop.Owing to its role in sustainable agriculture as both a rotation and a cash crop,its global market is expanding and increased production is urgently needed.For both technical and regulatory reasons,neither conventional nor transgenic breeding techniques can keep pace with the demand for increased production.In answer to this challenge,CRISPR/Cas9 genome editing technology has been gaining traction in plant biology and crop breeding in recent years.However,there are currently no reports of the successful application of the CRISPR/Cas9 genome editing technology in pea.We developed a transient transformation system of hairy roots,mediated by Agrobacterium rhizogenes strain K599,to validate the efficiency of a CRISPR/Cas9 system.Further optimization resulted in an efficient vector,PsU6.3-tRNA-PsPDS3-en35S-PsCas9.We used this optimized CRISPR/Cas9 system to edit the pea phytoene desaturase(PsPDS)gene,causing albinism,by Agrobacterium-mediated genetic transformation.This is the first report of successful generation of gene-edited pea plants by this route.展开更多
Protoplast has been widely used in biotechnologies to circumvent the breeding obstacles in citrus, including long juvenility, polyembryony, and male/female sterility. The protoplast-based transient gene expression sys...Protoplast has been widely used in biotechnologies to circumvent the breeding obstacles in citrus, including long juvenility, polyembryony, and male/female sterility. The protoplast-based transient gene expression system is a powerful tool for gene functional characterization and CRISPR/Cas9 genome editing in higher plants, but it has not been widely used in citrus. In this study, the polyethylene glycol(PEG)-mediated method was optimized for citrus callus protoplast transfection, with an improved transfection efficiency of 68.4%. Consequently, the efficiency of protein subcellular localization assay was increased to 65.8%, through transient expression of the target gene in protoplasts that stably express the fluorescent organelle marker protein. The gene editing frequencies in citrus callus protoplasts reached 14.2% after transient expression of CRISPR/Cas9 constructs. We demonstrated that the intronic polycistronic tRNAgRNA(inPTG) genome editing construct was functional in both the protoplast transient expression system and epicotyl stable transformation system in citrus. With this optimized protoplast transient expression system, we improved the efficiency of protein subcellular localization assay and developed the genome editing system in callus protoplasts, which provides an approach for prompt test of CRISPR vectors.展开更多
Base editors of the Cas9 system have been widely used for precise nucleotide substitution in crops. In this study, Cas12a was applied to construct plant cytosine base editors(CBEs). The main elements of Cas12aCBEs wer...Base editors of the Cas9 system have been widely used for precise nucleotide substitution in crops. In this study, Cas12a was applied to construct plant cytosine base editors(CBEs). The main elements of Cas12aCBEs were engineered and their efficiency was evaluated in stably transformed rice cells. An optimized ttCas12a-hyA3Bctd editor, consisting of a LbCas12a variant carrying catalytic inactive D832A and temperature-tolerance D156R double mutations, a truncated human APOBEC3B deaminase, a human RAD51 single-stranded DNA-binding domain, and double copies of UGI, outperformed other Cas12aCBEs in base editing efficiency. In T0transgenic rice plants, ttCas12a-hyA3Bctd edited an average of42.01% and a maximum of 68.75% of lines at six genomic targets. A-to-G conversions were generated in rice by an adenine base editor with a similar architecture to the optimized CBE. Our results provide preliminary evidence for the feasibility of robust and efficient plant Cas12a base editing systems, which could be useful for precise crop breeding.展开更多
Gene deletion has been a valuable tool for unraveling the mysteries of molecular biology.Early approaches included gene trapping and gene targetting to disrupt or delete a gene randomly or at a specific location,respe...Gene deletion has been a valuable tool for unraveling the mysteries of molecular biology.Early approaches included gene trapping and gene targetting to disrupt or delete a gene randomly or at a specific location,respectively.Using these technologies in mouse embryos led to the generation of mouse knocko ut models and many scientific discoveries.The efficacy and specificity of these approaches have significantly increased with the advent of new technology such as cluste red regula rly inters paced short palindromic repeats for targetted gene deletion.However,several limitations including unwanted off-target gene deletion have hindered their widespread use in the field.Crerecombinase technology has provided additional capacity for cell-specific gene deletion.In this review,we provide a summary of currently available literature on the application of this system for targetted deletion of neuronal genes.This article has been constructed to provide some background info rmation for the new trainees on the mechanism and to provide necessary information for the design,and application of the Cre-recombinase system thro ugh reviewing the most f requent promoters that are currently available for genetic manipulation of neuro ns.We additionally will provide a summary of the latest technological developments that can be used for targeting neurons.This may also serve as a general guide for the selection of appropriate models for biomedical research.展开更多
The Directory of China’s Foreign Trade,compiled by China Council for the Promotionof International Trade (CCPIT) and ChinaChamber of International Commerce(CCOIC), is always well received by friendsfrom foreign trade...The Directory of China’s Foreign Trade,compiled by China Council for the Promotionof International Trade (CCPIT) and ChinaChamber of International Commerce(CCOIC), is always well received by friendsfrom foreign trade companies and industrialenterprises both at home and abroad, and reprinted for several times.展开更多
Acta Geologica Sinica(English Edition)is a bimonthly,SCI-indexed academic journal produced by the Geological Society of China.With the latest 2022 Impact Factor of 3.3,it has long been indexed by many international da...Acta Geologica Sinica(English Edition)is a bimonthly,SCI-indexed academic journal produced by the Geological Society of China.With the latest 2022 Impact Factor of 3.3,it has long been indexed by many international databases and websites,such as SCI,CA,BIG,etc.Internationally,the journal cooperates with John Wiley&Sons,Inc.to publish the electronic version;all papers can be downloaded online.Contributors wishing to submit,read,enquire and download can log in via http://mc.manuscriptcentral.com/ags.展开更多
文摘Acta Geologica Sinica(English Edition)is a bimonthly,SCI-indexed academic journal produced by the Geological Society of China.With the latest 2022 Impact Factor of 3.3,it has long been indexed by many international databases and websites,such as SCI,CA,BIG,etc.Internationally,the journal cooperates with John Wiley&Sons,Inc.to publish the electronic version;all papers can be downloaded online.Contributors wishing to submit,read,enquire and download can log in via http://mc.manuscriptcentral.com/ags.
文摘“Journal of Jilin University(Science Edition)”is a comprehensive academic journal in the fields of science sponsored by Jilin University and administrated by the Ministry of Education of the People's Republic of China.The journal started publication in 1955.The original name at starting publication was“Journal of Natural Science of Northeast People University”,which was changed into“Acta Scientiarum Naturalium Universitatis Jilinensis”in 1958 owing to the name change of the university.
文摘“Journal of Jilin University(Science Edition)” is a comprehensive academic journal in the fields of science sponsored by Jilin University and administrated by the Ministry of Education of the People's Republic of China.The journal started publication in 1955.The original name at starting publication was “Journal of Natural Science of Northeast People University”,which was changed into “Acta Scientiarum Naturalium Universitatis Jilinensis” in 1958 owing to the name change of the university.
文摘“Journal of Jilin University(Science Edition)”is a comprehensive academic journal in the fields of science sponsored by Jilin University and administrated by the Ministry of Education of the People's Republic of China.The journal started publication in 1955.The original name at starting publication was“Journal of Natural Science of Northeast People University”.
文摘“Journal of Jilin University(Science Edition)” is a comprehensive academic journal in the fields of science sponsored by Jilin University and administrated by the Ministry of Education of the People’s Republic of China.The journal started publication in 1955.The original name at starting publication was “Journal of Natural Science of Northeast People University”.
文摘“Journal of Jilin University(Science Edition)” is a comprehensive academic journal in the fields of science sponsored by Jilin University and administrated by the Ministry of Education of the People’s Republic of China.The journal started publication in 1955.The original name at starting publication was “Journal of Natural Science of Northeast People University”,which was changed into “Acta Scientiarum Naturalium Universitatis Jilinensis” in 1958 owing to the name change of the university.
基金supported by the National Natural Science Foundation of China (31771369)the Natural Science Foundation of Fujian, China (2023J01443)the China Agriculture Research System of the Ministry of Agriculture and MARA (CARS-16)
文摘Jute(Corchorus capsularis L.)is the second most important natural plant fiber source after cotton.However,developing an efficient gene editing system for jute remains a challenge.In this study,the transgenic hairy root system mediated by Agrobacterium rhizogenes strain K599 was developed for Meifeng 4,an elite jute variety widely cultivated in China.The transgenic hairy root system for jute was verified by subcellular localization and bimolecular fluorescence complementation(BiFC)assays.The CHLOROPLASTOS ALTERADOS 1(CcCLA1)gene,which is involved in the development of chloroplasts,was targeted for editing at two sites in Meifeng 4.Based on this hairy root transformation,the gRNA scaffold was placed under the control of cotton ubiquitin GhU6.7 and-GhU6.9 promoters,respectively,to assess the efficiency of gene editing.Results indicated the 50.0%(GhU6.7)and 38.5%(GhU6.9)editing events in the target 2 alleles(gRNA2),but no mutation was detected in the target 1 allele(gRNA1)in transgenic-positive hairy roots.CcCLA1 gene editing at gRNA2 under the control of GhU6.7 in Meifeng 4 was also carried out by Agrobacterium tumefaciens-mediated transformation.Two CcCLA1 mutants were albinic,with a gene editing efficiency of 5.3%.These findings confirm that the CRISPR/Cas9 system,incorporating promoter GhU6.7,can be used as a gene editing tool for jute.
基金supported by the National Natural Science Foundation of China(32001532 and 31860411)the National Key Research and Development Program of China,(2022YFF1000020)+1 种基金Hunan Seed Industry Innovation Project(2021NK1012)the Yunnan Tobacco Company Project(2020530000241009)。
文摘The development and maturation of the CRISPR/Cas genome editing system provides a valuable tool for plant functional genomics and genetic improvement.Currently available genome-editing tools have a limited number of targets,restricting their application in genetic research.In this study,we developed a novel CRISPR/Cas9 plant ultra-multiplex genome editing system consisting of two template vectors,eight donor vectors,four destination vectors,and one primer-design software package.By combining the advantages of Golden Gate cloning to assemble multiple repetitive fragments and Gateway recombination to assemble large fragments and by changing the structure of the amplicons used to assemble sg RNA expression cassettes,the plant ultra-multiplex genome editing system can assemble a single binary vector targeting more than 40 genomic loci.A rice knockout vector containing 49 sg RNA expression cassettes was assembled and a high co-editing efficiency was observed.This plant ultra-multiplex genome editing system advances synthetic biology and plant genetic engineering.
文摘Aims and Scope Journal of Northeast Agricultural University(English Edition)is a comprehensive academic journal on agricultural sciences sponsored by Northeast Agricultural University and distributed worldwide.It is a peer reviewed journal published quarterly and mainly publishes review and research articles that reflect the latest achievements on crop science,horticulture,plant protection,resource and environment,animal science,veterinary medicine,agricultural engineering and technology,agricultural water conservancy,life science,biotechnology and food science.
文摘Aims and Scope Journal of Northeast Agricultural University(English Edition) is a comprehensive academic journal on agricultural sciences sponsored by Northeast Agricultural University and distributed worldwide. It is a peer reviewed journal published quarterly and mainly publishes review and research articles that reflect the latest achievements on crop science, horticulture, plant protection, resource and environment, animal science, veterinary medicine, agricultural engineering and technology, agricultural water conservancy, life science, biotechnology and food science.
文摘Aims and Scope Journal of Northeast Agricultural Univer-sity(English Edition)is a comprehensive academic journal on agricultural sciences sponsored by Northeast Agricultural University and distributed worldwide.It is a peer reviewed journal published quarterly and mainly publishes review and research articles that reflect the latest achievements on crop science,horticulture,plant protection,resource and environment,animal science,veterinary medicine,agricultural engineering and technology,agricultural water conservancy,life science,biotechnology and food science.
基金grants from the Special Project of Ministry of Science and Technology(Grant No.2017 YFC0113104)We thank Dr.Jiuhong Chen for helpful comments,Dr.Fabienne Hirigoyenberry and Dr.Huijun Chen,for language editing and LinkDoc for their constructive advice in patient follow-up.
文摘Objective:To validate the eighth edition of the AJCC staging system in patients with pancreatic adenocarcinoma receiving only stereotactic body radiation therapy and chemotherapy,and to propose modifications to improve prognostic accuracy.Methods:Patients with pathologically confirmed pancreatic adenocarcinoma without metastasis who were undergoing only chemoradiotherapy were included and staged according to the seventh and eighth editions of the AJCC staging system.Meanwhile,another group of stage T4 patients from the above enrollment with only portal vein involvement with or without tumor thrombi(PV±PVTT)were retrieved for survival comparisons.Modifications were proposed according to the survival comparisons.A cohort from the SEER database was used for external validation of the modified staging system.Results:A total of 683 patients were included.Patients with N2 or N1 but different T stages had significantly different survival outcomes according to the eighth edition.The survival of patients with(PV±PVTT)was comparable to that of patients with T4 tumors.The concordance index of the seventh and eighth editions,and the modified staging system was 0.744(95%CI:0.718—0.769),0.750(95%CI:0.725—0.775),and 0.788(95%CI:0.762-0.813),respectively.For external validation,the concordance index was 0.744(95%CI:0.718-0.770),0.750(95%CI:0.724-0.776),and 0.788(95%CI:0.762-0.814),respectively.Conclusions:The modified staging system is suggested to have the m ost accurate prognostic value.Hence,PV土PVTT should be added to the definition of T4 tumors regardless of tumor size.Patients with N2 or N1 in different T stages could be regrouped into different substages.Additionally,stage III should be subclassified into IIIA(T3N 2 and T4N 0)and IIIB(T4N 1-2).
基金the financial support of the China Agriculture Research System of MOF and MARA-Food Legumes(CARS-08)the Agricultural Science and Technology Innovation Program(ASTIP)of the Chinese Academy of Agricultural Sciences。
文摘Pea(Pisum sativum L.)is an annual cool-season legume crop.Owing to its role in sustainable agriculture as both a rotation and a cash crop,its global market is expanding and increased production is urgently needed.For both technical and regulatory reasons,neither conventional nor transgenic breeding techniques can keep pace with the demand for increased production.In answer to this challenge,CRISPR/Cas9 genome editing technology has been gaining traction in plant biology and crop breeding in recent years.However,there are currently no reports of the successful application of the CRISPR/Cas9 genome editing technology in pea.We developed a transient transformation system of hairy roots,mediated by Agrobacterium rhizogenes strain K599,to validate the efficiency of a CRISPR/Cas9 system.Further optimization resulted in an efficient vector,PsU6.3-tRNA-PsPDS3-en35S-PsCas9.We used this optimized CRISPR/Cas9 system to edit the pea phytoene desaturase(PsPDS)gene,causing albinism,by Agrobacterium-mediated genetic transformation.This is the first report of successful generation of gene-edited pea plants by this route.
基金supported by the National Natural Science Foundation of ChinaChina (Grant Nos. 31872051, 32072528)the Foundation of Hubei Hongshan Laboratory (Grant No.2021hszd009)。
文摘Protoplast has been widely used in biotechnologies to circumvent the breeding obstacles in citrus, including long juvenility, polyembryony, and male/female sterility. The protoplast-based transient gene expression system is a powerful tool for gene functional characterization and CRISPR/Cas9 genome editing in higher plants, but it has not been widely used in citrus. In this study, the polyethylene glycol(PEG)-mediated method was optimized for citrus callus protoplast transfection, with an improved transfection efficiency of 68.4%. Consequently, the efficiency of protein subcellular localization assay was increased to 65.8%, through transient expression of the target gene in protoplasts that stably express the fluorescent organelle marker protein. The gene editing frequencies in citrus callus protoplasts reached 14.2% after transient expression of CRISPR/Cas9 constructs. We demonstrated that the intronic polycistronic tRNAgRNA(inPTG) genome editing construct was functional in both the protoplast transient expression system and epicotyl stable transformation system in citrus. With this optimized protoplast transient expression system, we improved the efficiency of protein subcellular localization assay and developed the genome editing system in callus protoplasts, which provides an approach for prompt test of CRISPR vectors.
基金funded by the National Natural Science Foundation of China (U19A2022 and 32000284)the Natural Science Foundation of Anhui Province (2208085Y11, 2108085Y07,2008085QC101, and 2008085MC71)+2 种基金the University Synergy Innovation Program of Anhui Province (GXXT-2021-056)Open Research Fund Program of Anhui Province Key Laboratory of Rice Genetics and Breeding (SDKF-2021-01 and SDKF-2022-04)Natural Science Research Project for Anhui Universities(KJ2021A0196)。
文摘Base editors of the Cas9 system have been widely used for precise nucleotide substitution in crops. In this study, Cas12a was applied to construct plant cytosine base editors(CBEs). The main elements of Cas12aCBEs were engineered and their efficiency was evaluated in stably transformed rice cells. An optimized ttCas12a-hyA3Bctd editor, consisting of a LbCas12a variant carrying catalytic inactive D832A and temperature-tolerance D156R double mutations, a truncated human APOBEC3B deaminase, a human RAD51 single-stranded DNA-binding domain, and double copies of UGI, outperformed other Cas12aCBEs in base editing efficiency. In T0transgenic rice plants, ttCas12a-hyA3Bctd edited an average of42.01% and a maximum of 68.75% of lines at six genomic targets. A-to-G conversions were generated in rice by an adenine base editor with a similar architecture to the optimized CBE. Our results provide preliminary evidence for the feasibility of robust and efficient plant Cas12a base editing systems, which could be useful for precise crop breeding.
文摘Gene deletion has been a valuable tool for unraveling the mysteries of molecular biology.Early approaches included gene trapping and gene targetting to disrupt or delete a gene randomly or at a specific location,respectively.Using these technologies in mouse embryos led to the generation of mouse knocko ut models and many scientific discoveries.The efficacy and specificity of these approaches have significantly increased with the advent of new technology such as cluste red regula rly inters paced short palindromic repeats for targetted gene deletion.However,several limitations including unwanted off-target gene deletion have hindered their widespread use in the field.Crerecombinase technology has provided additional capacity for cell-specific gene deletion.In this review,we provide a summary of currently available literature on the application of this system for targetted deletion of neuronal genes.This article has been constructed to provide some background info rmation for the new trainees on the mechanism and to provide necessary information for the design,and application of the Cre-recombinase system thro ugh reviewing the most f requent promoters that are currently available for genetic manipulation of neuro ns.We additionally will provide a summary of the latest technological developments that can be used for targeting neurons.This may also serve as a general guide for the selection of appropriate models for biomedical research.
文摘The Directory of China’s Foreign Trade,compiled by China Council for the Promotionof International Trade (CCPIT) and ChinaChamber of International Commerce(CCOIC), is always well received by friendsfrom foreign trade companies and industrialenterprises both at home and abroad, and reprinted for several times.
文摘Acta Geologica Sinica(English Edition)is a bimonthly,SCI-indexed academic journal produced by the Geological Society of China.With the latest 2022 Impact Factor of 3.3,it has long been indexed by many international databases and websites,such as SCI,CA,BIG,etc.Internationally,the journal cooperates with John Wiley&Sons,Inc.to publish the electronic version;all papers can be downloaded online.Contributors wishing to submit,read,enquire and download can log in via http://mc.manuscriptcentral.com/ags.