Design, execution, and analysis of pooled <em>in vitro em>CRISPR/Cas9 screens
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- 作者：Linde A. Miles ; Ralph J. Garippa ; John T. Poirier
- 刊名：FEBS Journal
- 出版年：2016
- 出版时间：September 2016
- 年：2016
- 卷：283
- 期：17
- 页码：3170-3180
- 全文大小：469.5KB
- ISSN：1742-4658
文摘
The recently described clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 technology has proven to be an exquisitely powerful and invaluable method of genetic manipulation and/or modification. As such, many researchers have realized the potential of using the CRISPR/Cas9 system as a novel screening method for the identification of important proteins in biological processes and have designed short guide RNA libraries for an <em>in vitroem> screening. The seminal papers describing these libraries offer valuable information regarding methods for generating the short guide RNA libraries, creating cell lines containing these libraries, and specific details regarding the screening workflow. However, certain considerations are often overlooked that may be important when planning and performing a screen, including which CRISPR library to use and how to best analyze the resulting screen data. In this review, we offer suggestions to answer some of these questions that are not covered as deeply in the papers describing the available CRISPR libraries for an <em>in vitroem> screening.