Section of Cell and Developmental Biology, University of California, San Diego, 9500 Gilman Drive, La Jolla, CA, 92093-0335, USA.
Tata Institute for Genetics and Society-India (TIGS), TIGS Center at inStem, Bangalore, 560065, India.
Nat Commun. 2019 Apr 9;10(1):1640. doi: 10.1038/s41467-019-09694-w.
Gene-drive systems developed in several organisms result in super-Mendelian inheritance of transgenic insertions. Here, we generalize this "active genetic" approach to preferentially transmit allelic variants (allelic-drive) resulting from only a single or a few nucleotide alterations. We test two configurations for allelic-drive: one, copy-cutting, in which a non-preferred allele is selectively targeted for Cas9/guide RNA (gRNA) cleavage, and a more general approach, copy-grafting, that permits selective inheritance of a desired allele located in close proximity to the gRNA cut site. We also characterize a phenomenon we refer to as lethal-mosaicism that dominantly eliminates NHEJ-induced mutations and favors inheritance of functional cleavage-resistant alleles. These two efficient allelic-drive methods, enhanced by lethal mosaicism and a trans-generational drive process we refer to as "shadow-drive", have broad practical applications in improving health and agriculture and greatly extend the active genetics toolbox.
几种生物中开发的基因驱动系统导致转基因插入的超孟德尔遗传。在这里，我们将这种“主动遗传”方法推广到仅优先传递由单个或少数核苷酸改变引起的等位基因变体（等位基因驱动）。我们测试了两种等位基因驱动的配置：一种是复制切割，其中非优选等位基因被 Cas9/向导 RNA (gRNA) 切割选择性靶向，另一种是更通用的方法，复制嫁接，它允许位于 gRNA 切割位点附近的所需等位基因的选择性遗传。我们还描述了一种我们称之为致死镶嵌现象的现象，它主要消除 NHEJ 诱导的突变，并有利于具有功能的抗切割等位基因的遗传。通过致死镶嵌和我们称之为“影子驱动”的跨代驱动过程增强的这两种有效的等位基因驱动方法，在改善健康和农业方面具有广泛的实际应用，并极大地扩展了主动遗传学工具包。
