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Title: Hybrid cytomegalovirus enhancer-H1 promoter-based plasmid and baculovirus vectors mediate effective RNA interference
Authors: Ong, S.T.
Li, F.
Du, J.
Tan, Y.W.
Wang, S. 
Issue Date: Dec-2005
Citation: Ong, S.T., Li, F., Du, J., Tan, Y.W., Wang, S. (2005-12). Hybrid cytomegalovirus enhancer-H1 promoter-based plasmid and baculovirus vectors mediate effective RNA interference. Human Gene Therapy 16 (12) : 1404-1412. ScholarBank@NUS Repository.
Abstract: Plasmid and viral vectors harboring an RNA polymerase (Pol) III promoter would be useful in achieving sustained cellular expression of short interfering RNA (siRNA) to inhibit disease-associated genes. Given that transcription machineries directed by certain Pol II and III promoters may use common factors, we investigated whether the enhancer of the Pol II cytomegalovirus (CMV) immediate-early promoter could improve the efficacy of RNA interference mediated by the Pol III H1 promoter. We constructed a hybrid promoter by appending the CMV enhancer 5′ to the H1 promoter. In the context of plasmid vectors, the hybrid promoter provided up to 50% greater inhibition of the expression of target genes than the unmodified H1 promoter and extended the silencing effect beyond that provided by the H1 promoter. Insect baculoviruses can infect a broad range of mammalian cell types. We constructed a baculovector expression cassette in which the synthesis of short hairpin RNA was under the control of the hybrid CMV enhancer-H1 promoter. This recombinant baculovirus vector was capable of suppressing expression of a target gene by 95% in cultured cells and by 82% in vivo in rat brain. These findings indicate that the hybrid CMV enhancer-H1 promoter can be used favorably for RNA interference. © Mary Ann Liebert, Inc.
Source Title: Human Gene Therapy
ISSN: 10430342
DOI: 10.1089/hum.2005.16.1404
Appears in Collections:Staff Publications

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