Month: February 2022

Piggy Bac mediated transgenesis

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Piggy Bac mediated transgenesis offers many advantages over traditional transgenics. It has a large cargo capacity (up to 200 kb) allowing insertion of large promoters and regulatory elements. The insertion is footprint free and does not leave behind vector sequences. The transgene is also inserted in the host genome in a precise manner as a single copy without any chromosomal rearrangement or deletion (chromothripsis) and therefore offers reliable and consistent transgene expression.

 

In the Piggy Bac approach, the donor template includes the transgene of interest flanked by inverted terminal repeat (ITR) sequences. The donor template is microinjected into 1-cell embryos along with mRNA encoding the transposase, which integrates the transgene into the host genome through a “cut and paste” mechanism.

 

The CMGM has generated several novel mouse models using this approach including cell specific ablation and conditional rescue humanized mouse models.

PiggyBac

PiggyBac-on-BAC illustration