用于自噬通量评估的 Dual HiBiT-GFP-LC3 慢病毒报告系统。

A Dual HiBiT-GFP-LC3 Lentiviral Reporter for Autophagy Flux Assessment.

机构信息

Cellular Tools, Genomics and Proteomics Core Facility, German Cancer Research Center (DKFZ), Heidelberg, Germany.

Institute for Cell Biology, University Hospital Essen, Essen, Germany.

出版信息

Methods Mol Biol. 2022;2445:75-98. doi: 10.1007/978-1-0716-2071-7_6.

DOI:10.1007/978-1-0716-2071-7_6

PMID:34972987

Abstract

Autophagy is an intracellular degradation process that maintains the cellular homeostasis and it is regulated in multiple ways, both in health and disease. Assessment of autophagic flux in cells is an important approach for understanding the function of autophagy in biological contexts. Here, we describe a new tool for the qualitative and quantitative determination of autophagic flux using a dual lentiviral reporter system that generates a fusion HiBiT-GFP-LC3B protein suitable for generating stable cell lines.

摘要

自噬是一种维持细胞内环境稳定的细胞内降解过程，它在健康和疾病中都受到多种方式的调节。评估细胞中的自噬流是了解自噬在生物背景下功能的重要方法。在这里，我们描述了一种使用双慢病毒报告系统定性和定量测定自噬流的新工具，该系统生成适合生成稳定细胞系的融合 HiBiT-GFP-LC3B 蛋白。