CD4 aptamer–RORγt shRNA chimera inhibits IL-17 synthesis by human CD4+ T cells
| Autor: | Pingfang Song, Yuan K. Chou, Roberto Meza-Romero, Cong Qiu Chu, Kentaro Yomogida, Gil Benedek, Xiaowei Zhang |
|---|---|
| Rok vydání: | 2014 |
| Předmět: |
CD4-Positive T-Lymphocytes
biology Cellular differentiation Interleukin-17 Biophysics Cell Biology Transfection Nuclear Receptor Subfamily 1 Group F Member 3 Biochemistry Molecular biology Article Cell Line Small hairpin RNA Chimera (genetics) RNA interference Cell culture CD4 Antigens Gene expression biology.protein Humans Th17 Cells RNA Small Interfering Molecular Biology Dicer |
| Zdroj: | Biochemical and Biophysical Research Communications. 452:1040-1045 |
| ISSN: | 0006-291X |
| DOI: | 10.1016/j.bbrc.2014.09.037 |
| Popis: | Cell type specific delivery of RNAi to T cells has remained to be a challenge. Here we describe an aptamer mediated delivery of shRNA to CD4(+) T cells targeting RORγt to suppress Th17 cells. A cDNA encoding CD4 aptamer and RORγt shRNA was constructed and the chimeric CD4 aptamer-RORγt shRNA (CD4-AshR-RORγt) was generated using in vitro T7 RNA transcription. 2'-F-dCTP and 2'-F-dUTP were incorporated into CD4-AshR-RORγt for RNase resistance. CD4-AshR-RORγt was specifically uptaken by CD4(+) Karpas 299 cells and primary human CD4(+) T cells. The RORγt shRNA moiety of CD4-AshR-RORγt chimera was cleaved and released by Dicer. Furthermore, CD4-AshR-RORγt suppressed RORγt gene expression in Karpas 299 cells and CD4(+) T cells and consequently inhibited Th17 cell differentiation and IL-17 production. These results demonstrate that aptamer-facilitated cell specific delivery of shRNA represents a novel approach for efficient RNAi delivery and is potentially to be developed for therapeutics targeting specific T cells subtypes. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect