Targeted delivery of anti-coxsackievirus siRNAs using ligand-conjugated packaging RNAs.
Coxsackievirus B3 (CVB3) is a common pathogen of myocarditis. We previously synthesized a siRNA targeting the CVB3 protease 2A (siRNA/2A) gene and achieved reduction of CVB3 replication by 92% in vitro. However, like other drugs under development, CVB3 siRNA faces a major challenge of targeted delivery. In this study, we investigated a novel approach to deliver CVB3 siRNAs to a specific cell population (e.g. HeLa cells containing folate receptor) using receptor ligand (folate)-linked packaging RNA (pRNA) from bacterial phage phi29. pRNA monomers can spontaneously form dimers and multimers under optimal conditions by base-pairing between their stem loops. By covalently linking a fluorescence-tag to folate, we delivered the conjugate specifically to HeLa cells without the need of transfection. We further demonstrated that pRNA covalently conjugated to siRNA/2A achieved an equivalent antiviral effect to that of the siRNA/2A alone. Finally, the drug targeted delivery was further evaluated by using pRNA monomers or dimers, which carried both the siRNA/2A and folate ligand and demonstrated that both of them strongly inhibited CVB3 replication. These data indicate that pRNA as a siRNA carrier can specifically deliver the drug to target cells via its ligand and specific receptor interaction and inhibit virus replication effectively.
Zhang
Huifang M
HM
Department of Pathology and Laboratory Medicine, University of British Columbia, Canada.
Su
Yue
Y
Guo
Songchuan
S
Yuan
Ji
J
Lim
Travis
T
Liu
Jing
J
Guo
Peixuan
P
Yang
Decheng
D
D000998:Antiviral Agents / Q000378:metabolism*; D017103:Bacillus Phages / Q000235:genetics; D001483:Base Sequence; D016503:Drug Delivery Systems / Q000379:methods*; D029822:Enterovirus B, Human / Q000187:drug effects*; D015316:Genetic Therapy / Q000379:methods*; D006367:HeLa Cells; D006801:Humans; D008954:Models, Biological; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D034741:RNA, Small Interfering / Q000378:metabolism*
D000998:Antiviral Agents; D034741:RNA, Small Interfering