医学分子生物学杂志2025，Vol.22Issue(4)：305-310,6.DOI:10.3870/j.issn.1672-8009.2025.04.001
胰管注射AAV8-gRNA实现胰岛β细胞Adra2a基因编辑
Pancreatic Ductal Infusion of gRNA Carried by Adeno-Associated Vi-ruses for β-Cell Adra2a Gene Editing
摘要
Abstract
Objective This study aims to utilize the CRISPR/Cas9 system to achieve β-cell-specific gene knockout in the pancreas through adeno-associated virus(AAV8)carrying guide RNA(gRNA)targeting the Adra2a gene.Methods RIP2(rat insulin Ⅱ promoter)-Cre;Cas9KI/+mice were used to achieve β-cell-specific gene editing.In this model,the RIP2 promot-er drives Cre recombinase expression selectively in pancreatic β cells,resulting in targeted activa-tion of Cas9 in these cells.To enable efficient editing of the Adra2a gene,adeno-associated virus se-rotype 8,a low-immunogenic vector,was used to deliver gene-specific guide RNAs.AAV8 particles carrying gRNAs targeting Adra2a were injected via the pancreatic duct,enabling precise gene knockout in β cells.Results Successful β-cell-specific gene knockout was achieved in the pancre-as,confirmed by tissue analysis,PCR,and protein expression analysis.GFP expression was ob-served in the pancreas,and the targeted gene was successfully disrupted in β cells.Conclusion The use of AAV carrying gRNA in the RIP2-Cre;Cas9KI/+mouse model provides an effective method for β-cell-specific gene knockout,offering a valuable tool for future studies of islet gene function.关键词
基因编辑/CRISPR-Cas9/腺相关病毒/β细胞Key words
gene editing/CRISPR-Cas9/adeno-associated virus/β-cells分类
医药卫生引用本文复制引用
张欣,汪欣欣,吕婷婷,韩晓,朱云霞..胰管注射AAV8-gRNA实现胰岛β细胞Adra2a基因编辑[J].医学分子生物学杂志,2025,22(4):305-310,6.基金项目
国家自然科学基金面上项目(No.82270844) This work was supported by a grant from the National Natural Science Foundation of China(No.82270844) （No.82270844）
