RNA 介导的转基因精确插入的实验考虑因素。

4区生物学 Q3 Biochemistry, Genetics and Molecular Biology Methods in enzymology Pub Date : 2024-01-01 Epub Date: 2024-09-10 DOI:10.1016/bs.mie.2024.07.007

Sarah M Palm, Briana Van Treeck, Kathleen Collins

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引用次数: 0

摘要

精确 RNA 介导的转基因插入（PRINT）是一种用于人类基因组特异性、安全性转基因补充的开创性方法，它利用真核逆转录蛋白并完全依赖于 RNA 的传递。在这里，我们概述了设计两种所需 RNA 的重要注意事项、生产这些 RNA 并将其转染到细胞的细节，以及成功添加转基因的读数。在整个过程中，我们还提供了一些提示和关键概念，以帮助人们普遍使用这种方法。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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Experimental considerations for precise RNA-mediated insertion of transgenes.

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.

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来源期刊

Methods in enzymology 生物-生化研究方法

CiteScore

2.90

自引率

0.00%

发文量

308

审稿时长

3-6 weeks

期刊介绍： The critically acclaimed laboratory standard for almost 50 years, Methods in Enzymology is one of the most highly respected publications in the field of biochemistry. Each volume is eagerly awaited, frequently consulted, and praised by researchers and reviewers alike. Now with over 500 volumes the series contains much material still relevant today and is truly an essential publication for researchers in all fields of life sciences, including microbiology, biochemistry, cancer research and genetics-just to name a few. Five of the 2013 Nobel Laureates have edited or contributed to volumes of MIE.