| RRC ID |
49262
|
| 著者 |
Morishita H, Kaizuka T, Hama Y, Mizushima N.
|
| タイトル |
A new probe to measure autophagic flux in vitro and in vivo.
|
| ジャーナル |
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.
|
| 巻・号 |
13(4)
|
| ページ |
757-758
|
| 公開日 |
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 |
9.77
|
| 引用数 |
11
|
|
WOS 分野
|
CELL BIOLOGY
|
| リソース情報 |
| 遺伝子材料 |
pMRX-IP-GFP-LC3-RFP-LC3 delta G (RDB14600)
pMRX-IP-GFP-LC3-RFP (RDB14601). |
