A simplified system for the effective expression and delivery of functional mature microRNAs in mammalian cells

• DOI: 10.1038/s41417-019-0113-y
• Publication Date: 2019-06-20T12:02:51Z
• Authors: Jiaming Fan; Yixiao Feng; Ruyi Zhang; Wenwen Zhang; Yi Shu; Zongyue Zeng; Shifeng Huang; Linghuan Zhang; Bo Huang; Di Wu; Bo Zhang; Xi Wang; Yan Lei; Zhenyu Ye; Ling Zhao; Daigui Cao; Lijuan Yang; Xian Chen; Bin Liu; William Wagstaff; Fang He; Xiaoxing Wu; Jing Zhang; Jennifer Moriatis...; Michael J. Lee; Rex C. Haydon; Hue H. Luu; Ailong Huang; Tong-Chuan He; Shujuan Yan
• Keywords: Limiting

MicroRNAs (miRNAs) are ~22 nucleotide noncoding RNAs that involved in virtually all aspects of cellular process as their deregulations associated with many pathological conditions. Mature miRNAs (mMIRs) generated through a series tightly-regulated nuclear and cytoplasmic processing events the transcribed primary, precursor mMIRs. Effective manipulations miRNA expression enable us to gain insights into functions explore potential therapeutic applications. Currently, overexpression is achieved by using chemically-synthesized mimics, or shRNA-like stem-loop vectors express primary miRNAs, which limited low transfection efficacy rate-limiting processing. To overcome processing, we developed novel strategy mMIRs driven converging U6/H1 dual promoters. As proof-of-concept study, constructed mMIR for hsa-miR-223 hsa-Let-7a-1, demonstrated expressed effectively silenced target gene expression, specifically suppressed reporter activity, significantly affected cell proliferation, similar respective miRNAs. Furthermore, these can be easily converted retroviral adenoviral vectors. Collectively, our simplified system should valuable tool study and/or deliver miRNA-based therapeutics.