The authors present a protocol for using ΦC31 and Bxb1 ntegrases simultaneously to facilitate cassette exchange at a predefined location. This approach permits greater control and accuracy over integration. They also present a general method for using polymerase chain reaction assays to verify that the desired cassette exchange occurred successfully.
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BigField GEG Tech's insight:
Genetic engineering of iPs invloving 3 site specific recombinases. PhiC31 integrase was used to mediate initial placement of a single copy of a reprogramming plasmid into the genome at a safe location. A second phage integrase, Bxb1, was used to place the full-length dystrophin coding sequence into the same location, and Cre resolvase was utilized to excise unwanted sequences.
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The authors present a protocol for using ΦC31 and Bxb1 ntegrases simultaneously to facilitate cassette exchange at a predefined location. This approach permits greater control and accuracy over integration. They also present a general method for using polymerase chain reaction assays to verify that the desired cassette exchange occurred successfully.
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