通过 CRISPR/Cas 方法对人细胞系中的干细胞基因 LGR4 进行敲除调控。

Modulation of a Stem Cell Gene: LGR4 Knockout in a Human Cell Line by CRISPR/Cas Method.

机构信息

Department of Oral and Maxillofacial Plastic Surgery, Martin Luther University Halle-Wittenberg, Halle, Germany.

出版信息

Methods Mol Biol. 2021;2269:255-268. doi: 10.1007/978-1-0716-1225-5_18.

Abstract

The modulation of gene expression is essential for the investigation of function or involved pathway of a single gene of interest, in particular in the developmental/stem cell biology. The temporary knock down of gene expression via siRNA is a well-established but with a residual expression connected modulation method. The chapter describes the complete knockout of a defined target and allows a comprehensive study of different gene like the stem cell gene LGR4 (Leucine-rich repeat-containing G-protein-coupled receptor 4) using the new developed CRISPR/Cas method (clustered regularly interspaced short palindromic repeats).

摘要

基因表达的调控对于研究单个感兴趣基因的功能或涉及的途径至关重要，特别是在发育/干细胞生物学中。通过 siRNA 进行基因表达的瞬时敲低是一种经过充分验证的方法，但存在与残留表达相关的调控问题。本章描述了一种明确靶基因的完全敲除方法，并使用新开发的 CRISPR/Cas 方法（成簇规律间隔短回文重复）允许对不同基因（如干细胞基因 LGR4（富含亮氨酸重复的 G 蛋白偶联受体 4））进行全面研究。