Involvement of an intracellular vesicular transport process in naked-sgRNA-mediated TRUE gene silencing.
Mol Med Rep
; 12(4): 6365-9, 2015 Oct.
Article
en En
| MEDLINE
| ID: mdl-26238202
ABSTRACT
tRNase ZL-utilizing efficacious gene silencing (TRUE gene silencing) is an RNA-mediated gene expression control technology with therapeutic potential. Recently, our group demonstrated that a heptamer, mh1 (Bcl2), targeting human Bcl-2 mRNA, can be taken up by cells without the use of any transfection reagents and can induce the apoptosis of leukemia cells. However, little is known regarding the mechanism of naked small guide (sg)RNA uptake by cultured cells. Therefore, in the present study the effects of various inhibitors on the induction of apoptosis by naked sgRNA treatment were investigated in order to identify the uptake pathway required for sgRNA function in cultured cells. Addition of the endocytosis inhibitors chlorpromazine, nystatin or methylßcyclodextrin together with naked effective sgRNA was unable to diminish the apoptosisinducing effects of naked sgRNA or the reduction in target mRNA, suggesting that functional uptake of sgRNA by cells is clathrin, caveolae and raftindependent. Next, chloroquine, an inhibitor of lysosome acidification, and brefeldin A, an inhibitor that blocks protein transport from the Golgi apparatus to the endoplasmic reticulum were administered. In the presence of these compounds, the apoptosisinducing effects of naked sgRNA were reduced. These results suggest that a vesicular transport process is involved in sgRNAmediated TRUE gene silencing. A greater understanding of how naked sgRNAs enter cells and how they reach their target RNAs may aid in the design of more specificallytargeted and potent sgRNA drugs.
Texto completo:
1
Colección:
01-internacional
Asunto principal:
ARN Guía de Kinetoplastida
/
Silenciador del Gen
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Endocitosis
Límite:
Humans
Idioma:
En
Revista:
Mol Med Rep
Año:
2015
Tipo del documento:
Article
País de afiliación:
Japón