利用 Prime Editing 在 中进行精确的基因组工程。
Precise genome engineering in using prime editing.
机构信息
Department of Genetics, Blavatnik Institute, Harvard Medical School, Boston, MA 02115;
Department of Genetics, Blavatnik Institute, Harvard Medical School, Boston, MA 02115.
出版信息
Proc Natl Acad Sci U S A. 2021 Jan 5;118(1). doi: 10.1073/pnas.2021996118.
Abstract
Precise genome editing is a valuable tool to study gene function in model organisms. Prime editing, a precise editing system developed in mammalian cells, does not require double-strand breaks or donor DNA and has low off-target effects. Here, we applied prime editing for the model organism and developed conditions for optimal editing. By expressing prime editing components in cultured cells or somatic cells of transgenic flies, we precisely introduce premature stop codons in three classical visible marker genes, , , and Furthermore, by restricting editing to germ cells, we demonstrate efficient germ-line transmission of a precise edit in to 36% of progeny. Our results suggest that prime editing is a useful system in to study gene function, such as engineering precise point mutations, deletions, or epitope tags.
摘要
精确的基因组编辑是研究模式生物中基因功能的一种有价值的工具。在哺乳动物细胞中开发的精确编辑系统 Prime Editing 不需要双链断裂或供体 DNA，并且脱靶效应较低。在这里，我们将 Prime Editing 应用于模式生物 ，并开发了最佳编辑条件。通过在培养细胞或转基因蝇的体细胞中表达 Prime Editing 组件，我们在三个经典的可见标记基因 、 、和 中精确地引入了过早终止密码子。此外，通过将编辑限制在生殖细胞中，我们证明了在 中精确编辑到 的高效种系传递，在 36%的后代中。我们的结果表明，Prime Editing 是一个在 中研究基因功能的有用系统，例如工程精确的点突变、缺失或表位标签。
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