Synthetic design of asymmetric miRNA with engineered 3′-overhang to improve strand selection

Kadekar, SandeepUppsala universitet,Polymerkemi,Translational Chemical Biology Laboratory (author)

We have developed a novel miRNA design that significantly improves strand selection within the RISC complex by engineering the 3′-end by adding extra nucleotides. Addition of seven nucleotides at the 3′-ends of the miR or miR* strand resulted in a thermodynamic asymmetry at either of the two-ends, which resulted in selective RISC recruitment as demonstrated by the stem-loop quantitative PCR experiment. Such selective recruitment was also corroborated at the protein level by Western blot analysis. In order to investigate the functional effect due to selective recruitment, we performed apoptosis and metastasis studies using human colon carcinoma cells (HCT116) and human osteosarcoma cells (MG63). These experiments indicated that the recruitment of miR strand is responsible for inducing apoptosis as well as to inhibit invasiveness of cancer cells. Recruitment of miR* strand, on the other hand, showed opposite effect. To the best of our knowledge, our strand engineering strategy is the first report of improved strand selection of desired miRNA strand by RISC without using any chemical modifications or mismatches. We believe such structural modifications of miR34a could mitigate some of the off-target effects of miRNA therapy and would also allow a better understanding of sequence-specific gene regulation. Such a design could also be adapted to other miRNA to enhance their therapeutic potential.

Nawale, Ganesh N.Uppsala universitet,Polymerkemi,Translational Chemical Biology Laboratory(Swepub:uu)ganna194 (author)
Karlsson, KiraUppsala universitet,Polymerkemi,Translational Chemical Biology Laboratory (author)
Ålander, CeciliaUppsala universitet,Polymerkemi,Translational Chemical Biology Laboratory(Swepub:uu)cecal375 (author)
Podiyan, Oommen,1977-Bioengineering and Nanomedicine Lab, Faculty of Biomedical Sciences and Engineering, Tampere University of Technology, and BioMediTech Institute, 33720, Tampere, Finland (author)
Varghese, Oommen P.,1977-Uppsala universitet,Polymerkemi,Translational Chemical Biology Laboratory(Swepub:uu)oopva021 (author)
Uppsala universitetPolymerkemi (creator_code:org_t)

By the author/editor: Kadekar, Sandeep; Nawale, Ganesh N ...; Karlsson, Kira; Ålander, Cecilia; Podiyan, Oommen, ...; Varghese, Oommen ...

About the subject

NATURAL SCIENCES: NATURAL SCIENCES; and Chemical Science ...; and Polymer Chemistr ...

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