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用于通过核酸酶诱导的突变使细胞富集的替代报告器。

Surrogate reporters for enrichment of cells with nuclease-induced mutations.

机构信息

Department of Chemistry, Seoul National University, Gwanak-gu, Seoul, South Korea.

出版信息

Nat Methods. 2011 Oct 9;8(11):941-3. doi: 10.1038/nmeth.1733.

Abstract

Zinc-finger nucleases (ZFNs) and TAL-effector nucleases (TALENs) are powerful tools for creating genetic modifications in eukaryotic cells and organisms. But wild-type and mutant cells that contain genetic modifications induced by these programmable nucleases are often phenotypically indistinguishable, hampering isolation of mutant cells. Here we show that transiently transfected episomal reporters encoding fluorescent proteins can be used as surrogate genes for the efficient enrichment of endogenous gene-modified cells by flow cytometry.

摘要

锌指核酸酶 (ZFNs) 和 TAL 效应物核酸酶 (TALENs) 是在真核细胞和生物中进行基因修饰的强大工具。但是,由这些可编程核酸酶诱导的含有遗传修饰的野生型和突变型细胞在表型上通常无法区分,这阻碍了突变型细胞的分离。在这里,我们表明,瞬时转染的含有荧光蛋白的附加体报告基因可用于通过流式细胞术有效富集内源性基因修饰的细胞。

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