Fatality in mice due to oversaturation of cellular microRNA/short hairpin RNA pathways
| Autor: | Theresa A. Storm, Catherine L. Jopling, Mark A. Kay, Patricia L. Marion, Dirk Grimm, Felix H. Salazar, Kusum Pandey, Konrad L. Streetz, Corrine R. Davis |
|---|---|
| Rok vydání: | 2006 |
| Předmět: |
Hepatitis B virus
Small RNA Multidisciplinary RNA Mice Transgenic Karyopherins Biology Survival Analysis Molecular biology Liver Regeneration Cell biology RNAi Therapeutics Viral vector Small hairpin RNA Mice MicroRNAs Liver RNA interference microRNA Animals Humans Nucleic Acid Conformation Gene silencing RNA Interference |
| Zdroj: | Nature. 441:537-541 |
| ISSN: | 1476-4687 0028-0836 |
| DOI: | 10.1038/nature04791 |
| Popis: | RNA interference (RNAi) is a universal and evolutionarily conserved phenomenon of post-transcriptional gene silencing by means of sequence-specific mRNA degradation, triggered by small double-stranded RNAs. Because this mechanism can be efficiently induced in vivo by expressing target-complementary short hairpin RNA (shRNA) from non-viral and viral vectors, RNAi is attractive for functional genomics and human therapeutics. Here we systematically investigate the long-term effects of sustained high-level shRNA expression in livers of adult mice. Robust shRNA expression in all the hepatocytes after intravenous infusion was achieved with an optimized shRNA delivery vector based on duplex-DNA-containing adeno-associated virus type 8 (AAV8). An evaluation of 49 distinct AAV/shRNA vectors, unique in length and sequence and directed against six targets, showed that 36 resulted in dose-dependent liver injury, with 23 ultimately causing death. Morbidity was associated with the downregulation of liver-derived microRNAs (miRNAs), indicating possible competition of the latter with shRNAs for limiting cellular factors required for the processing of various small RNAs. In vitro and in vivo shRNA transfection studies implied that one such factor, shared by the shRNA/miRNA pathways and readily saturated, is the nuclear karyopherin exportin-5. Our findings have fundamental consequences for future RNAi-based strategies in animals and humans, because controlling intracellular shRNA expression levels will be imperative. However, the risk of oversaturating endogenous small RNA pathways can be minimized by optimizing shRNA dose and sequence, as exemplified here by our report of persistent and therapeutic RNAi against human hepatitis B virus in vivo. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|