Transgenic Service

The combination of electroporation and Adeno-Associated Virus (AAV) for generating genetically modified mice, often referred to as AAV-EP or CRISPR-READI (CRISPR RNP electroporation and AAV donor infection), is an efficient, microinjection-free method used to create large-fragment knock-in mice. This technique addresses the limitations of standard electroporation by using AAV to deliver large DNA repair templates (up to ~4.7 kb), which are then integrated into the genome by CRISPR/Cas9 ribonucleoproteins (RNPs) introduced via electroporation. 

This technique involves injecting a foreign DNA sequence into the pronucleus of fertilized mouse egg cells (zygotes) that are still in the one-cell stage. The injected zygotes are then carried to term by foster mice. Some of the new-born mice have integrated the foreign DNA into their own genome.

This technique uses endonuclease-mediated, targeted genome editing. Cas9, guide RNA and, if desired, DNA (repair template) are injected into the cytoplasm of fertilized mouse oocytes (zygotes) and these are then transferred into foster mice. Some of the resulting offspring have a corresponding genome modification at the intended location.

A desired DNA sequence is introduced into mouse embryonic stem cells (ES cells) which are then injected into early mouse embryos (morulas or blastocysts). The cells with the foreign DNA integrate into the embryo after injection. In this way, chimeric mice with mutations in certain genes can be produced (e.g. “knock-out” mice). The effects observed in mice with such knock-out mutations allow conclusions to be drawn about the biological function that the silenced gene normally performs.