CD4 aptamer-ROR?t shRNA chimera inhibits IL-17 synthesis by human CD4(+) T cells.

ABSTRACT: 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?t to suppress Th17 cells. A cDNA encoding CD4 aptamer and ROR?t shRNA was constructed and the chimeric CD4 aptamer-ROR?t shRNA (CD4-AshR-ROR?t) was generated using in vitro T7 RNA transcription. 2'-F-dCTP and 2'-F-dUTP were incorporated into CD4-AshR-ROR?t for RNase resistance. CD4-AshR-ROR?t was specifically uptaken by CD4(+) Karpas 299 cells and primary human CD4(+) T cells. The ROR?t shRNA moiety of CD4-AshR-ROR?t chimera was cleaved and released by Dicer. Furthermore, CD4-AshR-ROR?t suppressed ROR?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.

SUBMITTER: Song P 

PROVIDER: S-EPMC4216182 | biostudies-literature | 2014 Oct

REPOSITORIES: biostudies-literature