Silencing SARS‐CoV Spike protein expression in cultured cells by RNA interference

Abstract

The severe acute respiratory syndrome (SARS) has been one of the most epidemic diseases threatening human health all over the world. Based on clinical studies, SARSCoV (the SARS-associated coronavirus), a novel coronavirus, is reported as the pathogen responsible for the disease. To date, no e¡ective and speci¢c therapeutic method can be used to treat patients su¡ering from SARS-CoV infection. RNA interference (RNAi) is a process by which the introduced small interfering RNA (siRNA) could cause the degradation of mRNA with identical sequence speci¢city. The RNAi methodology has been used as a tool to silence genes in cultured cells and in animals. Recently, this technique was employed in anti-virus infections in human immunode¢ciency virus and hepatitis C/B virus. In this study, RNAi technology has been applied to explore the possibility for prevention of SARS-CoV infection. We constructed speci¢c siRNAs targeting the S gene in SARS-CoV. We demonstrated that the siRNAs could e¡ectively and speci¢cally inhibit gene expression of Spike protein in SARS-CoV-infected cells. Our study provided evidence that RNAi could be a tool for inhibition of SARS-CoV. ß 2004 Published by Elsevier B.V. on behalf of the Federation of European Biochemical Societies.

Reconstituted influenza virus envelopes as an efficient carrier system for cellular delivery of small-interfering RNAs

“…Possible target diseases include respiratory infections like influenza infection itself and the severe acute respiratory syndrome for which siRNA-based therapeutic approaches are already under development. 21,[65][66][67][68][69][70][71] In summary, virosomes combine the advantages of synthetic delivery systems (well defined, safe) with the desired properties of viral delivery systems (targeting to cells, active translocation of siRNA into the cytoplasm). They may therefore be an attractive addition to the methods for in vivo administration of siRNA, especially where delivery of siRNA to the respiratory epithelium or to cells with Fc receptors like dendritic cells, macrophages, and NK cells is envisaged.…”

ZNF536, a Novel Zinc Finger Protein Specifically Expressed in the Brain, Negatively Regulates Neuron Differentiation by Repressing Retinoic Acid-Induced Gene Transcription

“…Total RNA from different tissues from adult mice or cell lines was extracted using TRIzol reagent (Invitrogen). Total RNA (10 g) from adult mouse tissues were fractionated on a 1% formaldehyde agarose gel and transferred to a nitrocellulose membrane for Northern blotting (37). A ZNF536 probe (3,196 to 4,026 bp from the CDS of the ZNF536 gene) was labeled with 32 P using a Prime-a-Gene kit (Promega).…”

RNA Interference Technologies and Therapeutics

“…Viral genes that are essential for virus replication and host genes that are essential for virus entry or that play an essential role in the virus life cycle constitute attractive targets. Silencing of genes relevant to viral infection, such as HIV, [128] hepatitis, [129] and severe acute respiratory syndrome (SARS)-associated coronavirus, [130] has been achieved using RNAi.…”