TALEN construction for porcine IκBα gene and the detection of knockout activity
DOI:
https://doi.org/10.56093/ijans.v87i8.73417Keywords:
IκBα gene, Porcine, Knockout, SSA, TALENAbstract
Xenotransplantation often causes severe immunological rejection. The IκBα (inhibitor of nuclear factor kappa B alpha) gene plays an important role in delayed immunological rejection. Research shows that IκBα mutations can mitigate delayed immunological rejection. In the present study, the first exon sequence of the porcine IκBα gene was used to construct a transcription activator-like effector nuclease (TALEN) plasmid pair, and we used the double-strand break repair via single strand annealing (SSA) method to examine the knockout efficiency of the TALEN plasmid pair. The results showed that the knockout activity was 3.955, and the difference was extremely significant compared with the control group. A TALEN plasmid pair targeting the first exon sequence of the IκBα gene, was successfully constructed for the first time, and this study lays the foundation for knocking out the porcine IκBα gene to mitigate the delayed immunological rejection induced by xenotransplantation.
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