Streamlined ex vivo and in vivo genome editing in mouse embryos using recombinant adeno-associated viruses.
Streamlined ex vivo and in vivo genome editing in mouse embryos using recombinant adeno-associated viruses.
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DOI：
10.1038/s41467-017-02706-7
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发表时间：
2018-01-29
影响因子：
16.6
通讯作者：
Rivera-Pérez JA
中科院分区：
文献类型：
--
作者：
Yoon Y;Wang D;Tai PWL;Riley J;Gao G;Rivera-Pérez JA
使用CRISPR-Cas9方法的最新进展显着提高了啮齿动物遗传操作的容易性。尽管如此，自30多年前最初描述小鼠转基因以来，将核酸递送到植入前胚胎中的方法几乎没有改变。在这里，我们报告了一种通过重组腺相关病毒（rAAV）将CRISPR-Cas9组分转导到植入前小鼠胚胎中来产生转基因小鼠的新策略。使用这种方法，我们有效地产生了各种有针对性的突变在胚胎中，包括插入缺失事件产生的非同源末端连接和定制的突变使用同源性定向修复。我们还通过将rAAV颗粒直接递送到怀孕雌性的输卵管中来实现体内基因修饰。我们的方法大大简化了转基因小鼠的产生，更重要的是，为其他哺乳动物物种的简化基因编辑打开了大门。CRISPR-Cas9已被广泛用于科学研究中的啮齿动物基因操作。在本文中，作者使用rAAV在外植体培养或体内培养中使用CRISPR-Cas9组件来培养小鼠胚胎，以产生基因编辑的动物。
Recent advances using CRISPR-Cas9 approaches have dramatically enhanced the ease for genetic manipulation in rodents. Notwithstanding, the methods to deliver nucleic acids into pre-implantation embryos have hardly changed since the original description of mouse transgenesis more than 30 years ago. Here we report a novel strategy to generate genetically modified mice by transduction of CRISPR-Cas9 components into pre-implantation mouse embryos via recombinant adeno-associated viruses (rAAVs). Using this approach, we efficiently generated a variety of targeted mutations in explanted embryos, including indel events produced by non-homologous end joining and tailored mutations using homology-directed repair. We also achieved gene modification in vivo by direct delivery of rAAV particles into the oviduct of pregnant females. Our approach greatly simplifies the generation of genetically modified mice and, more importantly, opens the door for streamlined gene editing in other mammalian species. CRISPR-Cas9 has been widely adopted for genetically manipulating rodents for scientific research. Here the authors transduce mouse embryos with CRISPR-Cas9 components using rAAVs in explant culture or in vivo to produce gene-edited animals.
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影响因子：
64.8
作者：
Komor AC;Kim YB;Packer MS;Zuris JA;Liu DR
通讯作者：
Liu DR
影响因子：
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Hsu PD;Lander ES;Zhang F
通讯作者：
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Dickinson ME;Flenniken AM;Ji X;Teboul L;Wong MD;White JK;Meehan TF;Weninger WJ;Westerberg H;Adissu H;Baker CN;Bower L;Brown JM;Caddle LB;Chiani F;Clary D;Cleak J;Daly MJ;Denegre JM;Doe B;Dolan ME;Edie SM;Fuchs H;Gailus-Durner V;Galli A;Gambadoro A;Gallegos J;Guo S;Horner NR;Hsu CW;Johnson SJ;Kalaga S;Keith LC;Lanoue L;Lawson TN;Lek M;Mark M;Marschall S;Mason J;McElwee ML;Newbigging S;Nutter LM;Peterson KA;Ramirez-Solis R;Rowland DJ;Ryder E;Samocha KE;Seavitt JR;Selloum M;Szoke-Kovacs Z;Tamura M;Trainor AG;Tudose I;Wakana S;Warren J;Wendling O;West DB;Wong L;Yoshiki A;International Mouse Phenotyping Consortium;Jackson Laboratory;Infrastructure Nationale PHENOMIN, Institut Clinique de la Souris (ICS);Charles River Laboratories;MRC Harwell;Toronto Centre for Phenogenomics;Wellcome Trust Sanger Institute;RIKEN BioResource Center;MacArthur DG;Tocchini-Valentini GP;Gao X;Flicek P;Bradley A;Skarnes WC;Justice MJ;Parkinson HE;Moore M;Wells S;Braun RE;Svenson KL;de Angelis MH;Herault Y;Mohun T;Mallon AM;Henkelman RM;Brown SD;Adams DJ;Lloyd KC;McKerlie C;Beaudet AL;Bućan M;Murray SA
通讯作者：
Murray SA
影响因子：
4.6
作者：
Hashimoto M;Takemoto T
通讯作者：
Takemoto T
影响因子：
56.9
作者：
Lois, C;Hong, EJ;Baltimore, D
通讯作者：
Baltimore, D