The authors used phiC31 integrase to insert a copy of a β-globin / puromycin (purr) vector into specific genomic locations of a human hematopoietic cell line. After genomic integration, they used Cre recombinase to remove the bacterial backbone and purr.Following that, the stable β-chain expression was continued for several months in the absence of selective pressure.
Proof of concept of how PhiC31 recombinase works in Capra hircus fibroblasts. Moreover, this study points out new pseudo attP sites in the mammalian genome.
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.
PLOS ONE: an inclusive, peer-reviewed, open-access resource from the PUBLIC LIBRARY OF SCIENCE. Reports of well-performed scientific studies from all disciplines freely available to the whole world.
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.
To get content containing either thought or leadership enter:
To get content containing both thought and leadership enter:
To get content containing the expression thought leadership enter:
You can enter several keywords and you can refine them whenever you want. Our suggestion engine uses more signals but entering a few keywords here will rapidly give you great content to curate.
The authors used phiC31 integrase to insert a copy of a β-globin / puromycin (purr) vector into specific genomic locations of a human hematopoietic cell line. After genomic integration, they used Cre recombinase to remove the bacterial backbone and purr.Following that, the stable β-chain expression was continued for several months in the absence of selective pressure.
www.geg-tech.com/Vectors