在1型糖尿病小鼠模型中，用于全基因组体内CRISPR筛选以研究自身免疫下β细胞保护作用的方案。

Protocol for genome-scale in vivo CRISPR screening to study protection of beta cells under autoimmunity in a type 1 diabetes mouse model.

作者信息

Li Jian, Lee Yu-Chi, Iessi Isabela L, Wu Chialing, Yi Peng, Cai Erica P

机构信息

Section for Islet Cell and Regenerative Biology, Joslin Diabetes Center, Harvard Medical School, Boston, MA 02215, USA.

Lilly Diabetes Center of Excellence, Indiana Biosciences Research Institute, Indianapolis, IN 46202, USA.

出版信息

STAR Protoc. 2023 Mar 13;4(2):102155. doi: 10.1016/j.xpro.2023.102155.

DOI:10.1016/j.xpro.2023.102155

PMID:36917606

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10025263/

Abstract

Autoimmunity-induced pancreatic beta cell failure is the main characteristic of type 1 diabetes (T1D). Here, we describe a protocol for genome-scale in vivo CRISPR-Cas9 screening for use in a mouse model of T1D. Using a non-obese-diabetic-derived mouse beta cell line, NIT-1, and a genome-wide CRISPR-Cas9 knockout library (GeCKO-v2), we describe how to identify genes that confer resistance to autoimmune killing. This protocol can be applied in other mouse models of autoimmunity. For complete details on the use and execution of this protocol, please refer to Cai et al. (2020)..

摘要

自身免疫诱导的胰腺β细胞功能衰竭是1型糖尿病（T1D）的主要特征。在此，我们描述了一种用于T1D小鼠模型的全基因组体内CRISPR-Cas9筛选方案。使用非肥胖糖尿病来源的小鼠β细胞系NIT-1和全基因组CRISPR-Cas9敲除文库（GeCKO-v2），我们描述了如何鉴定赋予抗自身免疫杀伤能力的基因。该方案可应用于其他自身免疫小鼠模型。有关本方案使用和执行的完整详细信息，请参考Cai等人（2020年）的文献。