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| 1 | Precise gene replacement in plants through CRISPR/Cas genome editing technology:current status and future perspectives显示文摘CRISPR/Cas,as a simple,versatile,robust and cost-effective system for genome manipulation,has dominated the genome editing field over the past few years.The application of CRISPR/Cas in crop improvement is particularly important in the context of global climate change,as well as diverse agricultural,environmental and ecological challenges.Various CRISPR/Cas toolboxes have been developed and allow for targeted mutagenesis at specific genome loci,transcriptome regulation and epigenome editing,base editing,and precise targeted gene/allele replacement or tagging in plants.In particular,precise replacement of an existing allele with an elite allele in a commercial variety through homology-directed repair(HDR)is a holy grail in genome editing for crop improvement as it has been very difficult,laborious and time-consuming to introgress the elite alleles into commercial varieties without any linkage drag from parental lines within a few generations in crop breeding practice.However,it still remains very challenging in crop plants.This review intends to provide an informative summary of the latest development and breakthroughs in gene replacement using CRISPR/Cas technology,with a focus on achievements,potential mechanisms and future perspectives in plant biological science as well as crop improvement. | Shaoya Li Lanqin Xia | 2020 | aBIOTECH2020，1，1： | 5 |
| 2 | 提高CRISPR/Cas9介导的动物基因组精确插入效率研究进展显示文摘基因编辑技术是指通过人为方式在基因组插入、缺失或替换特定碱基,对遗传物质进行精确修饰和定向编辑的一种技术。近年来,锌指核酸内切酶(zinc-finger endonuclease, ZFN)、类转录激活因子效应物核酸酶(transcription activator-like effector nuclease, TALEN)、成簇规律间隔短回文重复序列及其相关系统(clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9, CRISPR/Cas9)等基因编辑技术的出现,使特异性靶向修饰动物基因组序列成为可能。虽然利用CRISPR/Cas9等基因编辑工具可以在细胞基因组高效产生双链断裂(double-strand breaks, DSB),但利用同源定向修复(homology directed repair, HDR)介导的精确插入(knock in, KI)效率却十分低下。本文结合当前基因编辑技术的发展现状,对目前提高CRISPR/Cas9介导的动物基因组KI策略进行了综述,以期为人类疾病模型制备、基因治疗和家畜遗传改良等提供借鉴。 | 李国玲 杨善欣 吴珍芳 张献伟 | 2020 | 遗传2020，42，7： | 4 |
| 3 | Present and future prospects for wheat improvement through genome editing and advanced technologies显示文摘Wheat(Triticum aestivum,2n=6x=42,AABBDD)is one of the most important staple food crops in the world.Despite the fact that wheat production has significantly increased over the past decades,future wheat production will face unprecedented challenges from global climate change,increasing world population,and water shortages in arid and semi-arid lands.Furthermore,excessive applications of diverse fertilizers and pesticides are exacerbating environmental pollution and ecological deterioration.To ensure global food and ecosystem security,it is essential to enhance the resilience of wheat production while minimizing environmental pollution through the use of cutting-edge technologies.However,the hexaploid genome and gene redundancy complicate advances in genetic research and precision gene modifications for wheat improvement,thus impeding the breeding of elite wheat cultivars.In this review,we first introduce state-of-the-art genome-editing technologies in crop plants,especially wheat,for both functional genomics and genetic improvement.We then outline applications of other technologies,such as GWAS,high-throughput genotyping and phenotyping,speed breeding,and synthetic biology,in wheat.Finally,we discuss existing challenges in wheat genome editing and future prospects for precision gene modifications using advanced genome-editing technologies.We conclude that the combination of genome editing and other molecular breeding strategies will greatly facilitate genetic improvement ofwheat for sustainable global production. | Shaoya Li Chen Zhang Jingying Li Lei Yan Ning Wang Lanqin Xia | 2021 | Plant Communications2021，2，4： | 4 |
| 4 | Embryo-mediated genome editing for accelerated genetic improvement of livestock显示文摘Selecting beneficial DNA variants is the main goal of animal breeding.However,this process is inherently inefficient because each animal only carries a fraction of all desirable variants.Genome editing technology with its ability to directly introduce beneficial sequence variants offers new opportunities to modernize animal breeding by overcoming this biological limitation and accelerating genetic gains.To realize rapid genetic gain,precise edits need to be introduced into genomicallyselected embryos,which minimizes the genetic lag.However,embryo-mediated precision editing by homology-directed repair(HDR)mechanisms is currently an inefficient process that often produces mosaic embryos and greatly limits the numbers of available edited embryos.This review provides a summary of genome editing in bovine embryos and proposes an embryo-mediated accelerated breeding scheme that overcomes the present efficiency limitations of HDR editing in bovine embryos.It integrates embryo-based genomic selection with precise multi-editing and uses embryonic cloning with elite edited blastomeres or embryonic pluripotent stem cells to resolve mosaicism,enable multiplex editing and multiply rare elite genotypes.Such a breeding strategy would enable a more targeted,accelerated approach for livestock improvement that allows stacking of beneficial variants,even including novel traits from outside the breeding population,in the most recent elite genetic background,essentially within a single generation. | Zachariah MCLEAN Björn OBACK Götz LAIBLE | 2020 | Frontiers of Agricultural Science and Engineering2020，7，2： | 2 |
| 5 | Diverse Systems for Efficient Sequence Insertion and Replacement in Precise Plant Genome Editing显示文摘CRISPR-mediated genome editing has been widely applied in plants to make uncomplicated genomic modifications including gene knockout and base changes.However,the introduction of many genetic variants related to valuable agronomic traits requires complex and precise DNA changes.Different CRISPR systems have been developed to achieve efficient sequence insertion and replacement but with limited success.A recent study has significantly improved NHEJ-and HDR-mediated sequence insertion and replacement using chemically modified donor templates.Together with other newly developed precise editing systems,such as prime editing and CRISPR-associated transposases,these technologies will provide new avenues to further the plant genome editing field. | Yingxiao Zhang Yiping Qi | 2020 | BioDesign Research2020，，1： | 1 |
| 6 | 一种新型的CRISPR/Cas9-hLacI双链DNA供体适配基因编辑系统显示文摘在CRISPR/Cas9系统介导的基因编辑中,借助于双链DNA(double-stranded DNA,dsDNA)供体模板的重组效应能够实现对目标基因组靶位点的精确编辑和基因敲入,然而高等真核生物细胞中同源重组的低效性限制了该基因编辑策略的发展和应用。为提高CRISPR/Cas9系统介导dsDNA供体模板的同源重组效率,本研究利用大肠杆菌(Escherichia coli)乳糖操纵子阻遏蛋白LacI与操纵序列LacO特异性结合的特点,通过重组DNA技术将密码子人源化优化的阻遏蛋白基因LacI分别与脓链球菌(Streptococcus pyogenes)源的SpCas9和路邓葡萄球菌(Staphylococcus lugdunensis)源的SlugCas9-HF融合表达,通过PCR将操纵序列LacO与dsDNA供体嵌合,构建了新型的CRISPR/Cas9-hLacI供体适配系统(donor adapting system,DAS)。首先在报告载体水平上对Cas9核酸酶活性、DAS介导的同源引导修复(homology-directed repair,HDR)效率进行了验证和优化,其次在基因组水平对其介导的基因精确编辑进行了检测,并最终利用CRISPR/SlugCas9-hLacIDAS在HEK293T细胞中实现了VEGFA位点的精确编辑,效率高达30.5%,显著高于野生型。综上所述,本研究开发了新型的CRISPR/Cas9-hLacI供体适配基因编辑系统,丰富了CRISPR/Cas9基因编辑技术种类,为以后的基因编辑及分子设计育种研究提供了新的工具。 | 马宝霞 崔婕妤 钱泓润 张潇筠 杨森 张骐镜 韩艺帆 张智英 王建刚 徐坤 | 2023 | 生物工程学报2023，39，10： | 1 |
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