成年小鼠肝脏的快速体内多重编辑（RIME）。
Rapid in vivo multiplexed editing (RIME) of the adult mouse liver.
机构信息
Department of Medicine , University of Pennsylvania , Philadelphia , Pennsylvania , USA.
Department of Cell and Developmental Biology , University of Pennsylvania , Philadelphia , Pennsylvania , USA.
出版信息
Hepatology. 2023 Aug 1;78(2):486-502. doi: 10.1002/hep.32759. Epub 2022 Oct 13.
BACKGROUND AND AIMS
Assessing mammalian gene function in vivo has traditionally relied on manipulation of the mouse genome in embryonic stem cells or perizygotic embryos. These approaches are time-consuming and require extensive breeding when simultaneous mutations in multiple genes is desired. The aim of this study is to introduce a rapid in vivo multiplexed editing (RIME) method and provide proof of concept of this system.
APPROACH AND RESULTS
RIME, a system wherein CRISPR/caspase 9 technology, paired with adeno-associated viruses (AAVs), permits the inactivation of one or more genes in the adult mouse liver. The method is quick, requiring as little as 1 month from conceptualization to knockout, and highly efficient, enabling editing in >95% of target cells. To highlight its use, we used this system to inactivate, alone or in combination, genes with functions spanning metabolism, mitosis, mitochondrial maintenance, and cell proliferation.
CONCLUSIONS
RIME enables the rapid, efficient, and inexpensive analysis of multiple genes in the mouse liver in vivo .
背景与目的
传统上，评估哺乳动物基因在体内的功能依赖于在胚胎干细胞或围合期胚胎中对小鼠基因组的操作。这些方法耗时且需要大量繁殖，如果同时需要对多个基因进行突变。本研究的目的是引入一种快速体内多重编辑（RIME）方法，并提供该系统的概念验证。
方法和结果
RIME 是一种系统，其中 CRISPR/caspase 9 技术与腺相关病毒（AAVs）结合，可使成年小鼠肝脏中的一个或多个基因失活。该方法快速，从构思到敲除只需不到 1 个月的时间，且效率极高，可使>95%的靶细胞发生编辑。为了突出其用途，我们使用该系统单独或组合失活了具有代谢、有丝分裂、线粒体维持和细胞增殖功能的基因。
结论
RIME 能够快速、高效、廉价地在体内分析小鼠肝脏中的多个基因。
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