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Gene control using tRNA‐shRNA chimeras: knockdown by shRNA and degradation of tRNA (749.3)
Author(s) -
Zazueta Christopher,
Stork Devon,
Haushalter Karl
Publication year - 2014
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.28.1_supplement.749.3
Subject(s) - small hairpin rna , rna interference , gene knockdown , chimera (genetics) , transfer rna , rna , rna polymerase iii , gene , biology , microbiology and biotechnology , computational biology , chemistry , genetics , rna polymerase
Gene therapy using RNA interference (RNAi) mediated by short hairpin RNAs (shRNAs) targeted against HIV‐1 is a promising alternative to antiretroviral therapy. Fusing shRNA to carrier tRNA to produce a tRNA‐shRNA chimera can provide a tunable level of shRNA delivery that is dependent on the action of the enzyme tRNaseZ to separate the shRNA and tRNA components of the chimera. We have introduced changes in the sequence of the acceptor stem of the tRNA portion of the tRNA‐shRNA chimera to produce the next‐generation of tRNA‐shRNA chimeras. Our targeted mutations are designed to result in an isomerization that triggers rapid tRNA decay after release of the shRNA by tRNaseZ. The knockdown efficiency of the resulting constructs can be assayed by a dual luciferase assay. Grant Funding Source : Supported by NSF CHE‐1005253