RRC ID 49262
Author Morishita H, Kaizuka T, Hama Y, Mizushima N.
Title A new probe to measure autophagic flux in vitro and in vivo.
Journal Autophagy
Abstract Macroautophagy is a catabolic process that delivers cytoplasmic components via the autophagosome to lysosomes for degradation. Measuring autophagic activity is critical to dissect molecular mechanisms and functions of autophagy but remains challenging due to the lack of a definitive method. We have recently developed a new fluorescent probe, GFP-LC3-RFP-LC3ΔG, to assess autophagic flux. Upon intracellular expression, the probe is cleaved by ATG4 family proteases into equimolar amounts of GFP-LC3 and RFP-LC3ΔG. The former is degraded by autophagy while the latter persists as an internal control in the cytosol. Autophagic flux can thus be quantified by obtaining the ratio of GFP:RFP signals. Using this method, we have identified several autophagy-modulating drugs by screening an approved drug library. We have also demonstrated that induced and basal autophagic flux can be monitored in zebrafish and mice.
Volume 13(4)
Pages 757-758
Published 2017-4-3
DOI 10.1080/15548627.2016.1278094
PMID 28121224
PMC PMC5388228
MeSH Animals Autophagy* Green Fluorescent Proteins / metabolism Mice Microtubule-Associated Proteins / metabolism Molecular Probes / chemistry* Zebrafish
IF 11.059
Times Cited 11
WOS Category CELL BIOLOGY
Resource
DNA material pMRX-IP-GFP-LC3-RFP-LC3 delta G (RDB14600) pMRX-IP-GFP-LC3-RFP (RDB14601).