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CRISPR/Cas9: The Good, the Bad and the Ugly
March 30, 2017
3:00 PM CEST/9:00 AM EST
In the past years, CRISPR/Cas9 has shown to be a real revolution for generation of genetically engineered animals and a real hope for genetic therapy in humans. This powerful genome editing technology is working in a very large panel of species from plants to mammals. In rodents, numerous knock-out or point mutations models were easily generated with this approach. Among other advantages, it is fast (to obtain founder animals), quite easy (in comparison to ES route) and cost-effective. It also allows accessing a large panel of background and thus promise to be a powerful tool for polygenic studies.
In this session, we will present our results for generating complex mouse or rat models. We developed the CRISPR Mediated REarrangement (CRISMERE) strategy, which takes advantage of the CRISPR/Cas9 system, to generate easily most of the desired structural variant & CNV rearrangements. We were indeed able to achieve deletions, duplications, and inversions of genomic regions as large as 24.4 Mb in rat and mouse (the good).
We also used CRISPR/Cas9 to develop knock-in or conditional lines. We will show the actual limitations of this system for the modification of “big” genomic sequences in mouse and rat (the bad) and discuss possible solutions and explanations.
Finally, we will also show that very careful validation of the lines generated by CRISPR/Cas9 is requested (the ugly) and how some unexpected event can be used to generate targeted overexpressing models.
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