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Direct evidence of CRISPR-Cas9-mediated mitochondrial genome editing

The Innovation 2022年第6期3卷 126-136页

作者： Rui Bi Yu Li Min Xu Quanzhen Zheng Deng-Feng Zhang Xiao Li Guolan Ma Bolin Xiang Xiaojia Zhu Hui Zhao Xingxu Huang Ping Zheng Yong-Gang Yao Key Laboratory of Animal Models and Human Disease Mechanisms of the Chinese Academy of Sciences&Yunnan Province Kunming Institute of ZoologyChinese Academy of SciencesKunmingYunnan 650204China Kunming College of Life Science University of Chinese Academy of SciencesKunmingYunnan 650204China Center for Excellence in Brain Science and Intelligence Technology Chinese Academy of SciencesShanghai 200031China Kunming Biological Diversity Regional Center of Large Apparatus and Equipments Public Technology Service CenterKunming Institute of ZoologyChinese Academy of SciencesKunmingYunnan 650204China Key Laboratory for Regenerative Medicine Ministry of EducationSchool of Biomedical SciencesFaculty of MedicineChinese University of Hong Kongand Hong Kong Branch of CAS Center for Excellence in Animal Evolution and GeneticsHong Kong SARChina KIZ/CUHK Joint Laboratory of Bioresources and Molecular Research in Common Diseases Kunming Institute of ZoologyChinese Academy of SciencesKunmingYunnan 650204China School of Life Science and Technology ShanghaiTech UniversityShanghai 201210China State Key Laboratory of Genetic Resources and Evolution and Yunnan Key Laboratory of Animal ReproductionKunming Institute of ZoologyChinese Academy of SciencesKunmingYunnan 650204China Center for Excellence in Animal Evolution and Genetics Chinese Academy of SciencesKunmingYunnan 650204China

Pathogenic mitochondrial DNA(mtDNA)mutations can cause a variety of human *** recent development of genome-editing technologies to manipulate mtDNA,such as mitochondria-targeted DNA nucleases and base editors,offer a promising way for curing mitochondrial diseases caused by mtDNA *** CRISPR-Cas9 system is a widely used tool for genome editing;however,its application in mtDNA editing is still under *** this study,we developed a mito-Cas9 system by adding the mitochondria-targeted sequences and 30 untranslated region of nuclear-encoded mitochondrial genes upstream and downstream of the Cas9 gene,respectively.

关键词： targeted diseases editing

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CRISPR-Cas12a base editors confer efficient multiplexed genome editing in rice

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Plant Communications 2023年第4期4卷 8-11页

作者： Yanhao Cheng Yingxiao Zhang Gen Li Hong Fang Simon Sretenovic Avery Fan Jiang Li Jianping Xu Qiudeng Que Yiping Qi Department of Plant Science and Landscape Architecture University of MarylandCollege ParkMD 20742USA Seeds Research Syngenta Crop ProtectionLLC9 Davis DrResearch Triangle ParkNC 27709USA Syngenta Biotechnology China Ltd.25 Life Science Park Rd.Zhongguancun Life Science ParkBeijing 102206China Institute for Bioscience and Biotechnology Research University of MarylandRockvilleMD 20850USA

Dear Editor,Many Cas9-derived base editors have been developed for precise C-to-T and A-to-G base editing in plants(Molla et al.,2021).They are typically based on a SpCas9 nickase or its engineered variants with altered protospacer adjacent motif(PAM)requirements(Molla et al.,2021).CRISPR-Cas12a enables highly efficient multiplexed genome editing in plants,and its T-rich PAM preference complements the G-rich PAM requirement of SpCas9 in genome targeting(Zhang et al.,2019,2021).Because of the lack of an efficient Cas12a nickase,it has been challenging to develop efficient Cas12a base ***,Cas12a cytosine base editors(CBEs)and adenine base editors(ABEs)have been developed in mammalian cells(Li et al.,2018;Kleinstiver et al.,2019)with low DNA damage(Wang et al.,2020)because deactivated Cas12a(dCas12a)was ***,efficient dCas12a base editors are yet to be developed in plants.

关键词： base editing precise

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Preface to the special topic on gene editing towards translation

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National Science Open 2023年第5期2卷 3-4页

作者： Jinsong Li Anming Meng State Key Laboratory of Cell Biology CAS Center for Excellence in Molecular Cell ScienceShanghai Institute of Biochemistry and Cell BiologyChinese Academy of SciencesShanghai 200031China Laboratory of Molecular Developmental Biology State Key Laboratory of Membrane BiologyTsinghua-Peking Center for Life SciencesSchool of Life SciencesTsinghua UniversityBeijing 100084China

The first draft of the human genome,published in 2001,has had a broad effect on biomedical studies and *** the past two decades,the human genome project(HGP)has dramatically promoted our understanding of how genomic sequences work during development,diseases and ***,genome-editing technologies have been rapidly developed to precisely and efficiently rewrite DNA sequences of different species,especially in 2012 as the development of a revolutionary genome editing method based on clustered regularly interspaced short palindromic repeat(CRISPR)and CRISPR-associated(Cas)proteins.

关键词： evolution. sequences editing

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Efficient BoPDS Gene editing in Cabbage by the CRISPR/Cas9 System

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Horticultural Plant Journal 2019年第4期5卷 164-169页

作者： Cunfa Ma Mengci Liu Qinfei Li Jun Si Xuesong Ren Hongyuan Song Key Laboratory of Horticulture Science for Southern Mountains Regions Ministry of EducationCollege of Horticulture and Landscape ArchitectureSouthwest University

Genome editing offers great advantages in identifying gene function and generating agronomically important mutations in crops. Here,we report that the genome of cabbage(Brassica oleracea var. capitata), an important cruciferous vegetable, can also be precisely edited by a CRISPR/Cas9 system stacked with multiple single-guide RNA-expressing cassettes. When the phytoene desaturase Bo PDS gene was used as the target gene, an albino-phenotype transgenic shoot in T0 Basta-resistant lines was observed, and 37.5% of the transgenic cabbage shoots carried Bo PDS gene mutations as a result of nucleotide deletions at the expected position. Moreover, mutations were detected in sites with the same target sequence in gene Bol016089 which is paralogous to the Bo PDS gene. Our results show that the CRISPR/Cas9 system is a powerful tool for cabbage variety improvement by genome editing.

关键词： Cabbage CRISPR/Cas9 BoPDS Genome editing Mutation

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Precise genome editing without exogenous donor DNA via retron editing system in human cells

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Protein & Cell 2021年第11期12卷 899-902页

作者： Xiangfeng Kong Zikang Wang Renxia Zhang Xing Wang Yingsi Zhou Linyu Shi Hui Yang HUIGENE Therapeutics Inc. Shanghai 200131China Institute of Neuroscience State Key Laboratory of NeuroscienceKey Laboratory of Primate NeurobiologyCenter for Excellence in Brain Science and Intelligence TechnologyChinese Academy of SciencesShanghai 200031China College of Life Sciences University of Chinese Academy of SciencesBeijing 100049China Shanghai Center for Brain Science and Brain-Inspired Intelligence Technology Shanghai 201210China

Dear Editor,CRISPR-Cas9 mediated seamless genome editing can be achieved by incorporating donor DNA into the CRISPR-Cas9 target loci via homology-directed repair(HDR),albeit with relative low efficiency due to the inefficient delivery of exogenous DNA(Cox et al.,2015;Gao,2021).Retrons are bacterial phage-defense related operons composed of a specialized reverse transcriptase(RT)and a relevant non-coding RNA(ncRNA)which can be partially reverse tran-scribed by RT initiating at a conserved guanosine(G)residue to produce a multicopy single-stranded DNA(msDNA)(Yee et al.,1984;Millman et al.,2020).After being reverse transcribed,the msDNA is usually covalently teth-ered to the ncRNA through the 2',5'-phosphodiesterbond between the priming G in ncRNA and 5'end of msDNA(Dhundale et al.,1987).The reverse transcription process,of which the specialized RT recognizes the unique secondary structure of retron ncRNA,is highly specific(Hsu et al.,1989).Additionally,desired msDNA can be generated in vivo by replacing the dispensable region of retron ncRNA with desired sequences(Mirochnitchenko et al.,1994;Simon et al.,2019).Therefore,retrons are promising biological sources for in vivo generation of DNA donors for HDR-me-diated precise genome editing.

关键词： donor specialized editing

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Engineered exosomes for future gene-editing therapy

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Biomaterials Translational 2022年第4期3卷 240-242页

作者： Haoyu Guo Xin Huang Department of Orthopaedics Union HospitalTongji Medical CollegeHuazhong University of Science and TechnologyWuhanHubei ProvinceChina

The RNA-guided clustered,regularly interspaced,short palindromic repeats(CRISPR)-associated nuclease protein 9(Cas9)-based technology is an advanced and popular gene-editing technology,which has shown great potential in treating genetic disorders in both animal models and even in clinical trials.1,2 Many transportation strategies are available for delivery of the CRISPR-Cas9 system,among which Cas9 ribonucleoprotein(RNP)delivery has some competitive advantages over other options,such as faster editing onset speed with a reduced immune response and lower off-target activity.3,4 However,the low in vivo delivery efficiency and poor tissue specificity of RNP delivery have limited further clinical applications.

关键词： clinical specificity editing

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Harnessing the potential of gene editing technology using CRISPR in inflammatory bowel disease

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World Journal of Gastroenterology 2019年第18期25卷 2177-2187页

作者： Viktor Limanskiy Arpita Vyas Lakshmi Shankar Chaturvedi Dinesh Vyas Department of Surgery San Joaquin General Hospital College of Medicine CNSU College of Medicine and College of Pharmacy California Northstate University

The molecular scalpel of clustered regularly interspersed short palindromic repeats/CRISPR associated protein 9(CRISPR/Cas9) technology may be sharp enough to begin cutting the genes implicated in inflammatory bowel disease(IBD) and consequently decrease the 6.3 billion dollar annual financial healthcare burden in the treatment of IBD. For the past few years CRISPR technology has drastically revolutionized DNA engineering and biomedical research field. We are beginning to see its application in gene manipulation of sickle cell disease,human immunodeficiency virus resistant embryologic twin gene modification and IBD genes such as Gatm(Glycine amidinotransferase, mitochondrial),nucleotide-binding oligomerization domain-containing protein 2, KRT12 and other genes implicated in adaptive immune convergence pathways have been subjected to gene editing, however there are very few publications. Furthermore,since Crohn's disease and ulcerative colitis have shared disease susceptibility and share genetic gene profile, it is paramount and is more advantageous to use CRISPR technology to maximize impact. Although, currently CRISPR does have its limitations due to limited number of specific Cas enzymes, off-target activity,protospacer adjacent motifs and crossfire between different target sites. However,these limitations have given researchers further insight on how to augment and manipulate enzymes to enable precise gene excision and limit crossfire between target sites.

关键词： Clustered regularly interspersed short palindromic repeats Inflammatory bowel disease Crohn’s disease Ulcerative colitis Gene excision Gene editing Gene therapy Financial impact of inflammatory bowel disease on healthcare Clustered regularly interspersed short palindromic repeats crossfire

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Spectral editing experiment of solid state ^(13)C NMR spectra under dynamic nuclear polarization enhancement

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Chinese Science Bulletin 1996年第8期41卷 631-634页

作者：杨保联周建威胡红兵李丽云胡建治裘鉴卿叶朝辉 Laboratory of Magnetic Resonance and Atomic and Molecular Physics Wuhan Institute of Physics Chinese Academy of Sciences Wuhan 430071 China

13C NMR is a widely used NMR technique. But for some samples, such as polymerand biological macromolecule, it is very difficult to assign their 13C spectra sinee the peaksof these spectra are often seriously overlapping. In solution, a number of techniques areavailable fbr simplifying the complex spectra, such as ’distortionless enhaneement

关键词： solid state I3C-NMR spectral editing dynamic nuclear polarization.

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Synergistic engineering of CRISPR-Cas nucleases enables robust mammalian genome editing

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The Innovation 2022年第4期3卷 74-82页

作者： Yangcan Chen Yanping Hu Xinge Wang Shengqiu Luo Ning Yang Yi Chen Zhikun Li Qi Zhou Wei Li State Key Laboratory of Stem Cell and Reproductive Biology Institute of ZoologyChinese Academy of SciencesBeijing 100101China University of Chinese Academy of Sciences Beijing 100049China Institute for Stem Cell and Regenerative Medicine Chinese Academy of SciencesBeijing 100101China Bejing Institute for Stem Cell and Regenerative Medicine Beijing 100101China

The naturally occurring prokaryotic CRISPR-Cas systems provide valuable resources for the development of new genome-editing ***,the majority of prokaryotic Cas nucleases exhibit poor editing efficiency in mammalian cells,which significantly limits their ***,we have developed a method termed Improving editing Activity by Synergistic Engineering(MIDAS).This method exerts a synergistic effect to improve mammalian genome-editing efficiency of a wide range of CRISPR-Cas systems by enhancing the interactions of Cas nuclease with the protospacer adjacent motif(PAM)and the single-stranded DNA(ssDNA)substrate in the catalytic pocket *** robustly and significantly increased the gene-editing efficiency of Cas12i,Cas12b,and CasX in human ***,a Cas12i variant,Cas12iMax,exhibited robust activity with a very broad PAM range(NTNN,NNTN,NAAN,and NCAN)and higher efficiency than the current widely used Cas nucleases.A high-fidelity version of Cas12iMax(Cas12iHiFi)has been further engineered to minimize off-target *** work provides an expandable and efficacious method for engineering Cas nucleases for robust mammalian genome editing.

关键词： editing engineering mammalian

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Plant Genome editing Database (PGED): A Call for Submission of Information about Genome-Edited Plant Mutants

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Molecular Plant 2019年第2期12卷 127-129页

作者： Yi Zheng Ning Zhang Gregory B. Martin Zhangjun Fei Boyce Thompson Institute for Plant Research Ithaca NY 14853 USA Plant Pathology and Plant-Microbe Biology Section School of Integrative 2Plant Pathology and Plant-Microbe Biology Section School of Integrative Plant Science Cornell University Ithaca NY 14853 USA USDA-ARS Robert W. Holley Center for Agriculture and Health Ithaca NY 14853 USA These authors contributed equally to this article

Recent advances in genome editing technologies, particularly CRISPR/Cas, enable the alteration of DNA sequences to produce deletions, insertions, and substitutions in genes (Jaganathan et al., 2018), as well as large ... 详细信息

关键词： Plant Genome editing Database (PGED) Genome-Edited Plant Mutants

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