Intraperitoneal AAV9-shRNA inhibits target expression in neonatal skeletal and cardiac muscles

Systemic injections of AAV vectors generally transduce to the liver more effectively than to cardiac and skeletal muscles. The short hairpin RNA (shRNA)-expressing AAV9 (shRNA-AAV9) can also reduce target gene expression in the liver, but not enough in cardiac or skeletal muscles. Higher doses of shRNA-AAV9 required for inhibiting target genes in cardiac and skeletal muscles often results in shRNA-related toxicity including microRNA oversaturation that can induce fetal liver failure.In this study, we injected high-dose shRNA-AAV9 to neonates and efficiently silenced genes in cardiac and skeletal muscles without inducing liver toxicity. This is because AAV is most likely diluted or degraded in the liver than in cardiac or skeletal muscle during cell division after birth. We report that this systemically injected shRNA-AAV method does not induce any major side effects, such as liver dysfunction, and the dose of shRNA-AAV is sufficient for gene silencing in skeletal and cardiac muscle tissues. This novel method may be useful for generating gene knockdown in skeletal and cardiac mouse tissues, thus providing mouse models useful for analyzing diseases caused by loss-of-function of target genes.

Research highlights► RNAi is a powerful tool in molecular biology and gene therapy, but the delivery of siRNA/shRNA is a major obstacle for its in vivo application. ► This is a report that systemically injected shRNA-AAV inhibits the target gene in the skeletal and cardiac muscles. ► Moreover, this method clearly shows no side effects, especially liver dysfunction which is a major side effect of shRNA-AAV injection systemically.