targeted delivery of mutant tolerant anti-coxsackievirus artificial micrornas using folate conjugated bacteriophage phi29 prna目标交付突变宽容anti-coxsackievirus人工使用叶酸共轭小分子核糖核酸噬菌体phi29 prna.pdf

Targeted Delivery of Mutant Tolerant Anti- Coxsackievirus Artificial MicroRNAs Using Folate Conjugated Bacteriophage Phi29 pRNA Xin Ye, Zhen Liu, Maged Gomaa Hemida, Decheng Yang* Department of Pathology and Laboratory Medicine, The Institute for Heart and Lung Health, St. Paul’s Hospital, University of British Columbia, Vancouver, British Columbia, Canada Abstract Background: Myocarditis is the major heart disease in infants and young adults. It is very commonly caused by coxsackievirus B3 (CVB3) infection; however, no specific treatment or vaccine is available at present. RNA interference (RNAi)-based anti-viral therapy has shown potential to inhibit viral replication, but this strategy faces two major challenges; viral mutational escape from drug suppression and targeted delivery of the reagents to specific cell populations. Methodology/Principal Findings: In this study, we designed artificial microRNAs (AmiRs) targeting the 39untranslated region (39UTR) of CVB3 genome with mismatches to the central region of their targeting sites. Antiviral evaluation showed that AmiR-1 and AmiR-2 reduced CVB3 (Kandolf and CG strains) replication approximately 100-fold in both HeLa cells and HL-1 cardiomyoctes. To achieve specific delivery, we linked AmiRs to the folate-conjugated bacterial phage packaging RNA (pRNA) and delivered the complexes into HeLa cells, a folate receptor positive cancer cells widely used as an in vitro model for CVB3 infection, via folate-mediated specific internalization. We found that our designed pRNA-AmiRs conjugates were tolerable to target mutations and have great potential to suppress viral mutational escape with little effect on triggering interferon induction. Conclusion/Significance: This study provides important clues for designing AmiRs targeting the 3 9UT