Department of Molecular and Cell Biology, University of California at Berkeley, Berkeley, CA, United States.
Department of Molecular and Cell Biology, University of California at Berkeley, Berkeley, CA, United States.
Methods Enzymol. 2024;705:1-24. doi: 10.1016/bs.mie.2024.07.007. Epub 2024 Sep 10.
Precise RNA-mediated insertion of transgenes (PRINT) is a pioneering method for site-specific, safe-harbor transgene supplementation of the human genome that harnesses a eukaryotic retroelement protein and relies solely on the delivery of RNA. Here we outline important considerations in the design of the two required RNAs, details for the production and transfection of these RNAs to cells, and read-outs for successful transgene addition. Throughout, tips and key concepts are laid out to enable general use of this method.
精确的 RNA 介导的转基因插入(PRINT)是一种开创性的方法,用于在人类基因组中进行特定位置、安全的转基因补充,该方法利用真核反转录元件蛋白,仅依赖 RNA 的递送。在这里,我们概述了在设计这两种必需 RNA 时的重要考虑因素,这些 RNA 的生产和转染到细胞中的详细信息,以及成功添加转基因的读出方法。在整个过程中,我们提供了一些技巧和关键概念,以方便该方法的普遍使用。