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A MUTANT T7 RNA POLYMERASE THAT OPTIMIZES THE YIELD OF RNA PRODUCTION |
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Principal Investigator: M. DREYFUS
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TECHNICAL DESCRIPTION
This invention describes significant improvement of the T7 RNA polymerase technology for RNA production. It describes RNA polymerase mutants that are less sensitive than the wild-type enzyme to abortive cycling over the first nucleotides of the transcribed DNA. These mutations are located in the N-terminal region, which participates in nucleic acid binding but not in catalysis. The mutants alleviate an important constraint on T7 RNAP transcription, i.e. the use of initial sequences resembling those of T7 late genes to minimize abortive transcription and thus optimize productive transcription. By decreasing the amount of valuable precursors wasted in abortive cycling, the new technology improves the yield and rate of synthesis of productive transcripts and facilitates their purification. These advantages are particularly obvious when the desired RNA is relatively short and its initial sequence is imposed.
Ref. : Guillerez, J., Lopez, J.P., Proux, F., Launay, H., Dreyfus, M. (2005) Proc. Natl. Acad. Sci. USA,, published on april 14, 2005 10.1073/pnas.0407141102.
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