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Base editing technology

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Frontiers of Agricultural Science and Engineering 2020年第2期7卷 227-228页

作者： Yidi SUN erwei zuo Hui YANG Key Laboratory of Systems Biology Center for Excellence in Molecular Cell ScienceInstitute of Biochemistry and Cell BiologyShanghai Institutes for Biological SciencesUniversity of Chinese Academy of SciencesChinese Academy of SciencesShanghai 200031China Bio-Med Big Data Center Key Laboratory of Computational BiologyCAS-MPG Partner Institute for Computational BiologyShanghai Institute of Nutrition and HealthShanghai Institutes for Biological SciencesUniversity of Chinese Academy of SciencesChinese Academy of Sciences(CAS)Shanghai 200031China Shenzhen Branch Guangdong Labratory for Lingnan Modern AgricultureGenome Analysis Laboratory of the Ministry of AgricultureAgricultural Genomics Institute at ShenzhenChinese Academy of Agricultural SciencesShenzhen 518124China Institute of Neuroscience State Key Laboratory of NeuroscienceKey Laboratory of Primate NeurobiologyCenter for Excellence in Brain Science and Intelligence TechnologyShanghai Research Center for Brain Science and Brain-Inspired IntelligenceShanghai Institutes for Biological SciencesChinese Academy of SciencesShanghai 200031China

The largest class of known human pathogenic mutations is point mutation,also known as single nucleotide polymorphism(SNP)[1].Precise and efficient editing of these SNPs has been of great interest for the treatment of genetic *** editing is a genome editing method which can precisely induce point mutations on DNA or RNA at the target loci without generating double strand breaks[2–5].

关键词： editing generating precisely

Indiscriminate ssDNA cleavage activity of CRISPR-Cas12a induces no detectable off-target effects in mouse embryos

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Protein & Cell 2021年第9期12卷 741-745页

作者： Yu Wei Yingsi Zhou Yajing Liu Wenqin Ying Ruiming Lv Qimeng Zhao Haibo Zhou erwei zuo Yidi Sun Hui Yang Changyang Zhou Institute of Neuroscienee State Key Laboratory of NeuroscieneeKey Laboratory of Primate NeurobiologyCenter for Excellence in Brain Science and Intelligenee TechnologyChinese Academy of SciencesShanghai 200031China University of Chinese Academy of Sciences Beijing 100049China Shanghai Center for Brain Science and Brairvlnspired Intelligence Technology Shanghai 201210China Center for Animal Genomics Agricultural Genome Institute at ShenzhenChinese Academy of Agricultural SciencesShenzhen 518124China

Dear Editor,Newly discovered characteristics like"collateral effect"or trans-cleavage in CRISPR-Cas13 and CRISPR-Cas12 systems have enabled their usage in nucleic acid detection(Gootenberg et al.2017,2018;Chen et al.2018).The collateral RNA cleavage of Cas13a has been reported to be harmful for cell development(Wang et al.2019;Buchman et al.2020).As a representative gene editor of CRISPR-Cas12 system,CRISPR-Cas12a(Cpf1)holds great potential for therapeutic applications in the future(Zetsche et al.2015;Koo et al.2018;Campa et al.2019).

关键词： al. cleavage holds

The advancements,challenges,and future implications of the CRISPR/Cas9 system in swine research

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Journal of Genetics and Genomics 2021年第5期48卷 347-360页

作者： Jinfu Zhang Emmanuel M.Khazalwa Hussein M.Abkallo Yuan Zhou Xiongwei Nie Jinxue Ruan Changzhi Zhao Jieru Wang Jing Xu Xinyun Li Shuhong Zhao erwei zuo Lucilla Steinaa Shengsong Xie Key Laboratory of Agricultural Animal Genetics Breeding and Reproduction of Ministry of Education&Key Lab of Swine Genetics and Breeding of Ministry of Agriculture and Rural AffairsHuazhong Agricultural UniversityWuhan 430070PR China Animal and Human Health Program BiosciencesInternational Livestock Research Institute(ILRI)P.O.Box 30709Nairobi 00100Kenya Key Laboratory of Pig Molecular Quantitative Genetics of Anhui Academy of Agricultural Sciences Livestock and Poultry Epidemic Diseases Research Center of Anhui ProvinceAnhui Provincial Key Laboratory of Livestock and Poultry Product Safety EngineeringInstitute of Animal Husbandry and Veterinary MedicineAnhui Academy of Agricultural SciencesHefei 230031PR China The Cooperative Innovation Center for Sustainable Pig Production Huazhong Agricultural UniversityWuhan 430070PR China sLingnan Guangdong Laboratory of Modern Agriculture Genome Analysis Laboratory of the Ministry of AgricultureAgricultural Genomics Institute at ShenzhenChinese Academy of Agricultural SciencesShenzhen 518124PR China

Clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR-associated protein 9(CRISPR/Cas9)genome editing technology has dramatically influenced swine research by enabling the production of high-quality disease-resistant pig breeds,thus improving *** addition,CRISPR/Cas9 has been used extensively in pigs as one of the tools in biomedical *** this review,we present the advancements of the CRISPR/Cas9 system in swine research,such as animal breeding,vaccine development,xenotransplantation,and disease *** also highlight the current challenges and some potential applications of the CRISPR/Cas9 technologies.

关键词： CRISPR/Cas9 Swine Genome editing CRISPR screening Base editing

Human 8-cell embryos enable efficient induction of disease-preventive mutations without off-target effect by cytosine base editor

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Protein & Cell 2023年第6期14卷 416-432页

作者： Yinghui Wei Meiling Zhang Jing Hu Yingsi Zhou Mingxing Xue Jianhang Yin Yuanhua Liu Hu Feng Ling Zhou Zhifang Li Dongshuang Wang Zhiguo Zhang Yin Zhou Hongbin Liu Ning Yao erwei zuo Jiazhi Hu Yanzhi Du Wen Li Chunlong Xu Hui Yang Center for Reproductive Medicine International Peace Maternity and Child Health HospitalInnovative Research Team of High-level Local Universities in ShanghaiSchool of MedicineShanghai Jiao Tong UniversityShanghai 2000301.Center for Reproductive MedicineInternational Peace Maternity and Child Health HospitalInnovative Research Team of High-level Local Universities in ShanghaiSchool of MedicineShanghai Jiao Tong UniversityShanghai 200030China Institute of Neuroscience State Key Laboratory of NeuroscienceKey Laboratory of Primate NeurobiologyCAS Center for Excellence in Brain Science and Intelligence TechnologyShanghai Institutes for Biological SciencesChinese Academy of SciencesShanghai 200031China Lingang Laboratory Shanghai Research Center for Brain Science and Brain-Inspired Intelligence TechnologyShanghai 200031China Center for Reproductive Medicine Ren Ji HospitalSchool of MedicineShanghai Key Laboratory for Assisted Reproduction and Reproductive GeneticsShanghai Jiao Tong UniversityShanghai 200127China The MOE Key Laboratory of Cell Proliferation and Differentiation School of Life SciencesGenome Editing Research CenterPeking UniversityBeijing 100871China Center for Reproductive Medicine Shandong UniversityJinan 250012China Center for Reproductive Medicine Anhui Provincial Maternal and Child Health HospitalHefei 230001China Shenzhen Branch Guangdong Laboratory for Lingnan Modern AgricultureGenome Analysis Laboratory of the Ministry of AgricultureAgricultural Genomics Institute at ShenzhenChinese Academy of Agricultural SciencesShenzhen 518000China Reproductive Medicine Center Department of Obstetrics and Gynecologythe First Affiliated Hospital of Anhui Medical UniversityHefei 230022China

Approximately 140 million people worldwide are homozygous carriers of APOE4(ε4),a strong genetic risk factor for late onset familial and sporadic Alzheimer’s disease(AD),91%of whom will develop AD at earlier age than heterozygous carriers and *** to AD could be reduced by targeted editing of APOE4,but a technical basis for controlling the off-target effects of base editors is necessary to develop low-risk personalized gene ***,we first screened eight cytosine base editor variants at four injection stages(from 1-to 8-cell stage),and found that FNLS-YE1 variant in 8-cell embryos achieved the comparable base conversion rate(up to 100%)with the lowest bystander *** particular,80%of AD-susceptibleε4 allele copies were converted to the AD-neutralε3 allele in humanε4-carrying *** control measures combined with targeted deep sequencing,whole genome sequencing,and RNA sequencing showed no DNA or RNA off-target events in FNLS-YE1-treated human embryos or their derived stem ***,base editing with FNLS-YE1 showed no effects on embryo development to the blastocyst ***,we also demonstrated FNLS-YE1 could introduce known protective variants in human embryos to potentially reduce human susceptivity to systemic lupus erythematosus and familial *** study therefore suggests that base editing with FNLS-YE1 can efficiently and safely introduce known preventive variants in 8-cell human embryos,a potential approach for reducing human susceptibility to AD or other genetic diseases.

关键词： human embryo APOE4 disease-preventive mutations base editor

Disruption of splicing-regulatory elements using CRISPR/Cas9 to rescue spinal muscular atrophy in human iPSCs and mice

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National Science Review 2020年第1期7卷 92-101页

作者： Jin-Jing Li Xiang Lin Cheng Tang Ying-Qian Lu Xinde Hu erwei zuo He Li Wenqin Ying Yidi Sun Lu-Lu Lai Hai-Zhu Chen Xin-Xin Guo Qi-Jie Zhang Shuang Wu Changyang Zhou Xiaowen Shen Qifang Wang Min-Ting Lin Li-Xiang Ma Ning Wang Adrian R.Krainer Linyu Shi Hui Yang Wan-Jin Chen Department of Neurology and Institute of Neurology The First Affiliated Hospital of Fujian Medical University Fujian Key Laboratory of Molecular Neurology Fujian Medical University Institute of Neuroscience State Key Laboratory of Neuroscience Key Laboratory of Primate Neurobiology CAS Center for Excellence in Brain Science and Intelligence Technology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Key Lab of Computational Biology CAS-MPG Partner Institute for Computational Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Department of Anatomy Histology & Embryology Shanghai Medical College Fudan University Cold Spring Harbor Laboratory Cold Spring Harbor

We here report a genome-editing strategy to correct spinal muscular atrophy(SMA). Rather than directly targeting the pathogenic exonic mutations, our strategy employed Cas9 and guide-sg RNA for the targeted disruption of intronic splicing-regulatory elements. We disrupted intronic splicing silencers(ISSs, including ISS-N1 and ISS + 100) of survival motor neuron(SMN) 2, a key modifier gene of SMA, to enhance exon 7 inclusion and full-length SMN expression in SMA iPSCs. Survival of splicing-corrected iPSC-derived motor neurons was rescued with SMN restoration. Furthermore, co-injection of Cas9 mRNA from Streptococcus pyogenes(SpCas9) or Cas9 from Staphylococcus aureus(SaCas9) alongside their corresponding sgRNAs targeting ISS-N1 into zygotes rescued 56% and 100% of severe SMA transgenic mice(Smn-/-, SMN2tg/-).The median survival of the resulting mice was extended to >400 days. Collectively, our study provides proof-of-principle for a new strategy to therapeutically intervene in SMA and other RNA-splicing-related diseases.

关键词： spinal muscular atrophy SMN2 splicing-regulatory elements CRISPR/Cas9 germline correction

数字化管道系统中心线控制点误差分析及修正——以天津LNG输气干线工程为例

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当代化工研究 2018年第4期 67-68页

作者：左二伟高鹏中石化江苏油建工程有限公司江苏225000

数字化管道系统的不断完善为油气管道建设的快速、高水平发展提供了强有力的技术保障,而中心线控制点坐标的完整和准确是数字化管道建设的关键环节。本文以天津LNG输气干线工程为例,对中心线控制点坐标的偏差进行了分析,并提出了修正方法。

关键词：中心线控制点坐标误差修正

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基因编辑技术的风险和机遇

科学 2019年第6期71卷 25-28页

作者：孙怡迪左二伟杨辉中国科学院上海生命科学研究院生物化学与细胞生物学研究所上海200031 中国科学院上海生命科学研究院神经科学研究所上海200031

人类大多数遗传病与基因突变有关,基因编辑技朮因可对特定DNA片段上的目标基因进行“编辑”,而被用来精确修复疾病基因,也成为治疗遗传性罕见病的理想策略;但是基因编辑技朮的脱靶效应可能引起包括癌症在内的很多种副作用,科字家正通过... 详细信息

人类大多数遗传病与基因突变有关,基因编辑技朮因可对特定DNA片段上的目标基因进行“编辑”,而被用来精确修复疾病基因,也成为治疗遗传性罕见病的理想策略;但是基因编辑技朮的脱靶效应可能引起包括癌症在内的很多种副作用,科字家正通过引人突变优化编辑工具,尝试减少甚至消灭脱靶效应。

关键词：基因编辑 CRISPR/Cas9 编辑效率脱靶效应基因治疗

Bridgman growth and characterization of langanite(La3Ga5.5Nb0.5O14,LGN) single crystal

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Journal of Rare Earths 2025年第03期 584-590页

作者： Chengkai Ren Libin Yin Shuai Wang Weirong Chen Sheng Wang Kainan Xiong Xiaoniu Tu Ningzhong Bao Yanqing Zheng Jun Chen erwei Shi Shanghai Institute of Ceramics Chinese Academy of Sciences State Key Laboratory of Material-Oriented Chemical Enganeering College of Chemical EngineeringNanjing Tech University The State Key Discipline Laboratory of Wide Bandgap Semiconductor Technology School of MicroelectronicsXidian University College of Materials Science and Chemical Engineering Ningbo University Beijing Advanced Innovation Center for Materials Genome Engineering Department of Physical ChemistryUniversity of Science and Technology Beijing Hainan University

As a promising optical and piezoelectric crystal,efficient growth of LGN single crystal is crucial for its practical ***,a langanite(La3Ga5.5Nb0.5O14,LGN) crystal with high quality was successfully grown by the Bridgman method along the Z *** order to prepare high-purity polycrystalline precursors for LGN crystal growth,the sintering conditions of LGN polycrystalline precursors were studied in *** melting point of LGN was also measured to provide a reference for the crystal growth *** the [001] oriented wafer,the full width at half maximum（EWHM） value of the nigh-resolution X-ray diffraction（HRXRD） analysis is 38.52",demonstrating that the LGN crystal exhibits excellent crystalline *** addition,we also measured the thermal properties and transmission spectrum of the as-grown LGN *** is found that the absorption peak at 1.85 μm of the LGN crystal grown in air using the Bridgman method disappears compared with previous reports(grown in N2+（1-3） vol% O2atmosphere),which is attributed to the oxygen-enriched growth *** phenomenon also occurs in other LGS-type disordered *** is believed that these findings may expand the potential applications of LGS series crystals at 1.85 μm.

关键词：

基于CRISPR系统的基因编辑技术的研究进展

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福建医科大学学报 2021年第3期55卷 187-191页

作者：郑基坛(综述) 闫娜娜左二伟(审校) 福建医科大学附属第一医院神经内科福建省神经病学研究所福州350005 中国农业科学院深圳农业基因组研究所深圳518120

CRISPR(clustered regularly interspaced short palindromic repeats)系统,即成簇的、间隔规律的短回文重复序列。该系统广泛存在于40%的细菌及90%的古细菌中,主要用于应对外源DNA或者病毒入侵的免疫防御[1]。根据该特性,Jinek等[2]将t... 详细信息

CRISPR(clustered regularly interspaced short palindromic repeats)系统,即成簇的、间隔规律的短回文重复序列。该系统广泛存在于40%的细菌及90%的古细菌中,主要用于应对外源DNA或者病毒入侵的免疫防御[1]。根据该特性,Jinek等[2]将tracrRNA及crRNA整合形成单链引导RNA(small guide RNA,sgRNA),该sgRNA由5′端负责与靶序列互补配对的20个核苷酸及3′端参与Cas9蛋白识别的发卡结构组成,首次在体外实现对靶标序列的切割,为CRISPR/Cas基因编辑技术的广泛应用奠定了重要基础。

关键词： CRISPR 单碱基编辑技术引导基因编辑技术