16/10/2019 - 11:00 - Auditori PRBB

In vivo oviduct electroporation by GONAD, reducing your 3R footprint

Sesiones científicas

Ian Blackstock

Sonidel Ltd.

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This new technology is a clear example of advancement in the 3 Rs, with reduction of animals, refinement of its use with fewer surgeries and replacement of vasectomized males and pseudo pregnancy females. New technology to generate genetically edited animal models with CRISPR without the need to micro-inject embryo, and without having to re-implant them into pseudo-gestant females afterwards, for their gestation.

This new technique was published by  Masato Ohtsuka and his collaborators at Kagoshima University in Japan. Method called GONAD, initials of "Genome-editing via Oviductal Nucleic Acids Delivery", in which CRISPR reagents are injected into the oviduct of females crossed the previous day with the male, immediately an electric discharge is applied to the through electroporation tweezers and finally the same embryos, edited in situ, progress their gestation in the same female.

i-GONAD: a robust method for in situ germline genome engineering using CRISPR nucleases. Masato Ohtsuka, Masahiro Sato, Hiromi Miura, Shuji Takabayashi, Makoto Matsuyama, Takayuki Koyano, Naomi Arifin, Shingo Nakamura, Kentaa Wad and Channabasavaiah B. Gurumurthy. Genome Biology 2018 19:25 Published: 26 February 2018.

They also adapted the method for the in situ genetic editing of rat embryos, naturally naming the new method as r-GONAD, with the rat "r". Successful production of genome-edited rats by the rGONAD method. Kobayashi T, Namba M, Koyano T, Fukushima M, Sato M, Ohtsuka M, Matsuyama M. BMC Biotechnol. 2018 Apr 2;18(1):19.