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人工法off-line HDF对血液透析充分性和高同型半胱氨酸血症的影响
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作者 吴飞飞 罗来敏 +2 位作者 范燕娜 张桂玲 龚燕燕 《实用临床医学(江西)》 CAS 2024年第4期19-22,27,共5页
目的探究人工法off-line HDF对维持性血液透析(MHD)患者透析充分性、高同型半胱氨酸血症(HHcy)的影响。方法随机选取2021至2023年于南昌大学第一附属高新医院血液透析中心稳定透析6个月以上的MHD患者30例,进行单中心自身对照研究,时长6... 目的探究人工法off-line HDF对维持性血液透析(MHD)患者透析充分性、高同型半胱氨酸血症(HHcy)的影响。方法随机选取2021至2023年于南昌大学第一附属高新医院血液透析中心稳定透析6个月以上的MHD患者30例,进行单中心自身对照研究,时长6个月。前3个月维持每周2次高通量血液透析(HFHD)联合1次on-line HDF,后3个月维持每周2次人工法off-line HDF联合1次on-line HDF。于研究第3个月、第6个月时用全自动生化分析仪检测透析前后血尿素氮(BUN)、血肌酐(SCr)、β2-微球蛋白(β2MG)、同型半胱氨酸(Hcy)、白蛋白(Alb)、C反应蛋白(CRP)、血红蛋白(Hb)、甲状旁腺素(iPTH)、磷(P)、钾(K)水平;计算血清尿素清除指数(Kt/V)、尿素下降率(URR)和β2MG清除率。结果人工法off-line HDF模式透析的Kt/V、URR及β2MG清除率均较显著高于HFHD模式透析(P<0.05);人工法off-lineHDF模式透析Kt/V、URR及β2MG清除率达到透析充分的百分率分别为83.33%、83.33%和93.33%,均显著高于HFHD模式透析(P<0.05)。与HFHD模式透析时相比,人工法off-lineHDF模式透析结束后,患者透析前Hcy水平降低,Alb水平升高(P<0.05)。结论人工法off-line HDF模式透析可显著增加Kt/V、URR和β2MG清除率,提高MHD患者的透析充分性达标率,且对HHcy具有改善作用。 展开更多
关键词 人工法off-line HDF 血液透析 透析充分性 高同型半胱氨酸血症 血液透析滤过
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Impact of Sedation Protocols on Elderly Patients Undergoing Mechanical Ventilation and Off-Line Weaning
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作者 Yihui Li Yamin Yuan +1 位作者 Jinquan Zhou Li Ma 《Journal of Clinical and Nursing Research》 2024年第4期322-333,共12页
The proportion of elderly patients in intensive care is increasing, and a significant proportion of them require mechanical ventilation. How to implement safe and effective mechanical ventilation for elderly patients,... The proportion of elderly patients in intensive care is increasing, and a significant proportion of them require mechanical ventilation. How to implement safe and effective mechanical ventilation for elderly patients, and when appropriate off-line is an important issue in the field of critical care medicine. Appropriate sedation can improve patient outcomes, but excessive sedation may lead to prolonged mechanical ventilation and increase the risk of complications. Elderly patients should be closely monitored and evaluated on an individual basis while offline, and the sedation regimen should be dynamically adjusted. This requires the healthcare team to consider the patient’s sedation needs, disease status, and pharmacodynamics and pharmacokinetics of the drug to arrive at the best strategy. Although the current research has provided valuable insights and strategies for sedation and off-line management, there are still many problems to be further explored and solved. 展开更多
关键词 Elderly patients Mechanical ventilation off-line strategy Sedation treatment
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Genome-edited rabbits:Unleashing the potential of a promising experimental animal model across diverse diseases 被引量:1
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作者 Yang Han Jiale Zhou +3 位作者 Renquan Zhang Yuru Liang Liangxue Lai Zhanjun Li 《Zoological Research》 SCIE CSCD 2024年第2期253-262,共10页
Animal models are extensively used in all aspects of biomedical research,with substantial contributions to our understanding of diseases,the development of pharmaceuticals,and the exploration of gene functions.The fie... Animal models are extensively used in all aspects of biomedical research,with substantial contributions to our understanding of diseases,the development of pharmaceuticals,and the exploration of gene functions.The field of genome modification in rabbits has progressed slowly.However,recent advancements,particularly in CRISPR/Cas9-related technologies,have catalyzed the successful development of various genome-edited rabbit models to mimic diverse diseases,including cardiovascular disorders,immunodeficiencies,agingrelated ailments,neurological diseases,and ophthalmic pathologies.These models hold great promise in advancing biomedical research due to their closer physiological and biochemical resemblance to humans compared to mice.This review aims to summarize the novel gene-editing approaches currently available for rabbits and present the applications and prospects of such models in biomedicine,underscoring their impact and future potential in translational medicine. 展开更多
关键词 Genome editing Animal model RABBIT CRISPR/Cas9 Genetic diseases
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Development and Therapeutic Applications of Precise Gene Editing Technology
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作者 ZHANG Yi-Meng YANG Xiao +1 位作者 WANG Jian LI Zhen-Hua 《生物化学与生物物理进展》 SCIE CAS CSCD 北大核心 2024年第10期2637-2647,共11页
The advent of gene editing represents one of the most transformative breakthroughs in life science,making genome manipulation more accessible than ever before.While traditional CRISPR/Cas-based gene editing,which invo... The advent of gene editing represents one of the most transformative breakthroughs in life science,making genome manipulation more accessible than ever before.While traditional CRISPR/Cas-based gene editing,which involves double-strand DNA breaks(DSBs),excels at gene disruption,it is less effective for accurate gene modification.The limitation arises because DSBs are primarily repaired via non-homologous end joining(NHEJ),which tends to introduce indels at the break site.While homology directed repair(HDR)can achieve precise editing when a donor DNA template is provided,the reliance on DSBs often results in unintended genome damage.HDR is restricted to specific cell cycle phases,limiting its application.Currently,gene editing has evolved to unprecedented levels of precision without relying on DSB and HDR.The development of innovative systems,such as base editing,prime editing,and CRISPR-associated transposases(CASTs),now allow for precise editing ranging from single nucleotides to large DNA fragments.Base editors(BEs)enable the direct conversion of one nucleotide to another,and prime editors(PEs)further expand gene editing capabilities by allowing for the insertion,deletion,or alteration of small DNA fragments.The CAST system,a recent innovation,allows for the precise insertion of large DNA fragments at specific genomic locations.In recent years,the optimization of these precise gene editing tools has led to significant improvements in editing efficiency,specificity,and versatility,with advancements such as the creation of base editors for nucleotide transversions,enhanced prime editing systems for more efficient and precise modifications,and refined CAST systems for targeted large DNA insertions,expanding the range of applications for these tools.Concurrently,these advances are complemented by significant improvements in in vivo delivery methods,which have paved the way for therapeutic application of precise gene editing tools.Effective delivery systems are critical for the success of gene therapies,and recent developments in both viral and non-viral vectors have improved the efficiency and safety of gene editing.For instance,adeno-associated viruses(AAVs)are widely used due to their high transfection efficiency and low immunogenicity,though challenges such as limited cargo capacity and potential for immune responses remain.Non-viral delivery systems,including lipid nanoparticles(LNPs),offer an alternative with lower immunogenicity and higher payload capacity,although their transfection efficiency can be lower.The therapeutic potential of these precise gene editing technologies is vast,particularly in treating genetic disorders.Preclinical studies have demonstrated the effectiveness of base editing in correcting genetic mutations responsible for diseases such as cardiomyopathy,liver disease,and hereditary hearing loss.These technologies promise to treat symptoms and potentially cure the underlying genetic causes of these conditions.Meanwhile,challenges remain,such as optimizing the safety and specificity of gene editing tools,improving delivery systems,and overcoming off-target effects,all of which are critical for their successful application in clinical settings.In summary,the continuous evolution of precise gene editing technologies,combined with advancements in delivery systems,is driving the field toward new therapeutic applications that can potentially transform the treatment of genetic disorders by targeting their root causes. 展开更多
关键词 precise gene editing CRISPR/Cas system base editing prime editing gene therapy
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Acta Geologica Sinica(English Edition)Calls for Submission of Review Papers
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《Acta Geologica Sinica(English Edition)》 SCIE CAS CSCD 2024年第5期1418-1418,共1页
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
关键词 PAPERS editION journal
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Germline Gene-Editing Creates Enhanced Livestock-Technical and Especially Ethical Issues Challenge Its Use in Humans
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作者 Jennifer Welsh 《Engineering》 SCIE EI CAS CSCD 2024年第2期3-5,共3页
Using clustered regularly interspaced short palindromic repeats(CRISPR)-based molecular tools,scientists are engineering-as they are also doing with plants.-animals with advantageous traits,like disease resistance and... Using clustered regularly interspaced short palindromic repeats(CRISPR)-based molecular tools,scientists are engineering-as they are also doing with plants.-animals with advantageous traits,like disease resistance and improved food yield.While these innovative techniques could one day be applied in humans,technical hurdles and ethical concerns likely place this possibility far in the future,The enhancements rely on germline gene editing,which alters the genes in a way that passes the changes on to offspring.Ger m-line gene editing differs from the somatic cell gene editing used in the highly promising new treatment recently approved for the human disease sickle cell anemia. 展开更多
关键词 LIKELY CREATE editING
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Brief Introduction to“Journal of Jilin University(Science Edition)”
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《吉林大学学报(理学版)》 CAS 北大核心 2024年第6期F0003-F0003,共1页
“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
关键词 edition) journal NORTHEAST
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Brief Introduction to “Journal of Jilin University(Science Edition)”
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《吉林大学学报(理学版)》 CAS 北大核心 2024年第5期F0003-F0003,共1页
“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
关键词 editION JOURNAL NORTHEAST
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FROM THE EDITOR-IN-CHIEF Innovative technologies significantly propel the development of casting industry
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作者 Yan-chun Lou 《China Foundry》 SCIE EI CAS CSCD 2024年第5期I0001-I0001,共1页
At this memorable moment,we are excited to celebrate the 20th anniversary of CHINA FOUNDRY journal.Since its inception in August 2004,this academic platform dedicated to disseminating Chinese casting technology has tr... At this memorable moment,we are excited to celebrate the 20th anniversary of CHINA FOUNDRY journal.Since its inception in August 2004,this academic platform dedicated to disseminating Chinese casting technology has traversed two full decades.Over this period,we have received strong support from numerous domestic and international industry experts and scholars. 展开更多
关键词 journal edit INDUSTRY
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Brief Introduction to“Journal of Jilin University(Science Edition)”
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《吉林大学学报(理学版)》 CAS 北大核心 2024年第3期F0003-F0003,共1页
“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
关键词 editION journal NORTHEAST
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Brief Introduction to “Journal of Jilin University (Science Edition)”
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《吉林大学学报(理学版)》 CAS 北大核心 2024年第1期F0003-F0003,共1页
“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
关键词 editION JOURNAL NORTHEAST
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Brief Introduction to “Journal of Jilin University(Science Edition)”
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《吉林大学学报(理学版)》 CAS 北大核心 2024年第2期F0003-F0003,共1页
“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
关键词 editION JOURNAL NORTHEAST
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Development of an Agrobacterium-mediated CRISPR/Cas9 gene editing system in jute(Corchorus capsularis)
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作者 Shaolian Jiang Qin Li +9 位作者 Xiangxue Meng Mengxin Huang Jiayu Yao Chuanyu Wang Pingping Fang Aifen Tao Jiantang Xu Jianmin Qi Shuangxia Jin Liwu Zhang 《The Crop Journal》 SCIE CSCD 2024年第4期1266-1270,共5页
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
关键词 JUTE Agrobacterium-mediated transformation Genome editing Hairy root system
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Journal of Northeast Agricultural University(English Edition) Instruction to Authors
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《Journal of Northeast Agricultural University(English Edition)》 CAS 2024年第2期F0003-F0003,共1页
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
关键词 editION NORTHEAST JOURNAL
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A simple and efficient CRISPR/Cas9 system permits ultra-multiplex genome editing in plants
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作者 Suting Wu Htin Kyaw +11 位作者 Zhijun Tong Yirong Yang Zhiwei Wang Liying Zhang Lihua Deng Zhiguo Zhang Bingguang Xiao William Paul Quick Tiegang Lu Guoying Xiao Guannan Qin Xue'an Cui 《The Crop Journal》 SCIE CSCD 2024年第2期569-582,共14页
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
关键词 CRISPR/Cas9 Multiplex genome editing Assembly system PLANT
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Journal of Northeast Agricultural University(English Edition) Instruction to Authors
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《Journal of Northeast Agricultural University(English Edition)》 CAS 2024年第1期F0003-F0003,共1页
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
关键词 editION NORTHEAST JOURNAL
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Journal of Northeast Agricultural University(English Edition) Instruction to Authors
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《Journal of Northeast Agricultural University(English Edition)》 CAS 2024年第3期F0003-F0003,共1页
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
关键词 editION NORTHEAST JOURNAL
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Engineering high amylose and resistant starch in maize by CRISPR/Cas9-mediated editing of starch branching enzymes
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作者 Mingzheng Ma Shanqiu Sun +5 位作者 Jinjie Zhu Xiantao Qi Gaoke Li Jianguang Hu Chuanxiao Xie Changlin Liu 《The Crop Journal》 SCIE CSCD 2024年第4期1252-1258,共7页
To improve the amylose content(AC)and resistant starch content(RSC)of maize kernel starch,we employed the CRISPR/Cas9 system to create mutants of starch branching enzyme I(SBEI)and starch branching enzyme IIb(SBEIIb).... To improve the amylose content(AC)and resistant starch content(RSC)of maize kernel starch,we employed the CRISPR/Cas9 system to create mutants of starch branching enzyme I(SBEI)and starch branching enzyme IIb(SBEIIb).A frameshift mutation in SBEI(E1,a nucleotide insertion in exon 6)led to plants with higher RSC(1.07%),lower hundred-kernel weight(HKW,24.71±0.14 g),and lower plant height(PH,218.50±9.42 cm)compared to the wild type(WT).Like the WT,E1 kernel starch had irregular,polygonal shapes with sharp edges.A frameshift mutation in SBEIIb(E2,a four-nucleotide deletion in exon 8)led to higher AC(53.48%)and higher RSC(26.93%)than that for the WT.E2 kernel starch was significantly different from the WT regarding granule morphology,chain length distribution pattern,X-ray diffraction pattern,and thermal characteristics;the starch granules were more irregular in shape and comprised typical B-type crystals.Mutating SBEI and SBEIIb(E12)had a synergistic effect on RSC,HKW,PH,starch properties,and starch biosynthesis-associated gene expression.SBEIIa,SS1,SSIIa,SSIIIa,and SSIIIb were upregulated in E12 endosperm compared to WT endosperm.This study lays the foundation for rapidly improving the starch properties of elite maize lines. 展开更多
关键词 MAIZE Gene editing Starch branching enzyme I Starch branching enzyme IIb
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Metabolic engineering and genome editing strategies for enhanced lipid production in microalgae
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作者 ANJANI DEVI CHINTAGUNTA SAMUDRALA PRASHANT JEEVAN KUMAR NUNE SATYA SAMPATH KUMAR 《BIOCELL》 SCIE 2024年第8期1181-1195,共15页
Depleting global petroleum reserves and skyrocketing prices coupled with succinct supply have been a grave concern,which needs alternative sources to conventional fuels.Oleaginous microalgae have been explored for enh... Depleting global petroleum reserves and skyrocketing prices coupled with succinct supply have been a grave concern,which needs alternative sources to conventional fuels.Oleaginous microalgae have been explored for enhanced lipid production,leading towards biodiesel production.These microalgae have short life cycles,require less labor,and space,and are easy to scale up.Triacylglycerol,the primary source of lipids needed to produce biodiesel,is accumulated by most microalgae.The article focuses on different types of oleaginous microalgae,which can be used as a feedstock to produce biodiesel.Lipid biosynthesis in microalgae occurs through fatty acid synthesis and TAG synthesis approaches.In-depth discussions are held regarding other efficient methods for enhancing fatty acid and TAG synthesis,regulating TAG biosynthesis bypass methods,blocking competing pathways,multigene approach,and genome editing.The most potential targets for gene transformation are hypothesized to be a malic enzyme and diacylglycerol acyltransferase while lowering phosphoenolpyruvate carboxylase activity is reported to be advantageous for lipid synthesis. 展开更多
关键词 Oleaginous microalgae BIODIESEL TAG synthesis Metabolic engineering Genome editing
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Development of a single transcript CRISPR/Cas9 toolkit for efficient genome editing in autotetraploid alfalfa
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作者 Haixia Zhao Siyi Zhao +12 位作者 Yingping Cao Xiping Jiang Lijuan Zhao Zhimeng Li Mengqi Wang Ruijuan Yang Chuanen Zhou Zhaoming Wang Feng Yuan Dongmei Ma Hao Lin Wenwen Liu Chunxiang Fu 《The Crop Journal》 SCIE CSCD 2024年第3期788-795,共8页
Alfalfa(Medicago sativa.L.)is a globally significant autotetraploid legume forage crop.However,despite its importance,establishing efficient gene editing systems for cultivated alfalfa remains a formidable challenge.I... Alfalfa(Medicago sativa.L.)is a globally significant autotetraploid legume forage crop.However,despite its importance,establishing efficient gene editing systems for cultivated alfalfa remains a formidable challenge.In this study,we pioneered the development of a highly effective ultrasonic-assisted leaf disc transformation system for Gongnong 1 alfalfa,a variety widely cultivated in Northeast China.Subsequently,we created a single transcript CRISPR/Cas9(CRISPR_2.0)toolkit,incorporating multiplex gRNAs,designed for gene editing in Gongnong 1.Both Cas9 and gRNA scaffolds were under the control of the Arabidopsis ubiquitin-10 promoter,a widely employed polymeraseⅡconstitutive promoter known for strong transgene expression in dicots.To assess the toolkit’s efficiency,we targeted PALM1,a gene associated with a recognizable multifoliate phenotype.Utilizing the CRISPR_2.0 toolkit,we directed PALM1 editing at two sites in the wild-type Gongnong 1.Results indicated a 35.1%occurrence of editing events all in target 2 alleles,while no mutations were detected at target 1 in the transgenic-positive lines.To explore more efficient sgRNAs,we developed a rapid,reliable screening system based on Agrobacterium rhizogenes-mediated hairy root transformation,incorporating the visible reporter MtLAP1.This screening system demonstrated that most purple visible hairy roots underwent gene editing.Notably,sgRNA3,with an 83.0%editing efficiency,was selected using the visible hairy root system.As anticipated,tetra-allelic homozygous palm1 mutations exhibited a clear multifoliate phenotype.These palm1 lines demonstrated an average crude protein yield increase of 21.5%compared to trifoliolate alfalfa.Our findings highlight the modified CRISPR_2.0 system as a highly efficient and robust gene editing tool for autotetraploid alfalfa. 展开更多
关键词 ALFALFA Gene editing CRISPR_2.0 toolkit Hairy root system Tetra-allelic homozygous mutants
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