화학공학소재연구정보센터
Biochemical and Biophysical Research Communications, Vol.452, No.4, 1040-1045, 2014
CD4 aptamer-ROR gamma t shRNA chimera inhibits IL-17 synthesis by human CD4(+) T cells
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 ROR gamma t to suppress Th17 cells. A cDNA encoding CD4 aptamer and ROR gamma t shRNA was constructed and the chimeric CD4 aptamer-ROR gamma t shRNA (CD4-AshR-ROR gamma t) was generated using in vitro T7 RNA transcription. 2'-F-dCTP and 2'-F-dUTP were incorporated into CD4-AshR-ROR gamma t for RNase resistance. CD4-AshR-ROR gamma t was specifically uptaken by CD4(+) Karpas 299 cells and primary human CD4(+) T cells. The ROR gamma t shRNA moiety of CD4-AshR-ROR gamma t chimera was cleaved and released by Dicer. Furthermore, CD4-AshR-ROR gamma t suppressed ROR gamma t 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. Published by Elsevier Inc.