利用生长素诱导的降解结构域在秀丽隐杆线虫中靶向内源性蛋白进行时空敲低。

Targeting endogenous proteins for spatial and temporal knockdown using auxin-inducible degron in Caenorhabditis elegans.

机构信息

Department of Zoology, The University of British Columbia, Vancouver, BC V6T 1Z3, Canada; Graduate Program in Cell & Developmental Biology, The University of British Columbia, Vancouver, BC V6T 1Z3, Canada; The Life Sciences Institute, The University of British Columbia, Vancouver, BC V6T 1Z3, Canada.

出版信息

STAR Protoc. 2023 Mar 17;4(1):102028. doi: 10.1016/j.xpro.2022.102028. Epub 2023 Jan 13.

DOI:10.1016/j.xpro.2022.102028

PMID:36640369

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9860162/

Abstract

The auxin-inducible degron (AID) provides reversible, spatiotemporal control for the knockdown of target proteins. Here, we present a protocol for AID-mediated protein knockdown in Caenorhabditis elegans. We describe steps for generating the knock-in mutants using two CRISPR-Cas9 genome editing techniques and preparing the auxin-containing nematode growth media (NGM) plates. We also detail AID-mediated spatiotemporal protein knockdown. For complete details on the use and execution of this protocol, please refer to Kurashina et al. (2021)..

摘要

生长素诱导的降解结构域（AID）为靶蛋白的敲低提供了可逆的、时空控制。在这里，我们提供了一个在秀丽隐杆线虫中进行 AID 介导的蛋白敲低的方案。我们描述了使用两种 CRISPR-Cas9 基因组编辑技术生成敲入突变体的步骤，以及准备含有生长素的线虫生长培养基（NGM）平板。我们还详细介绍了 AID 介导的时空蛋白敲低。有关此方案的使用和执行的完整详细信息，请参阅 Kurashina 等人。（2021）。