TY  - JOUR
AU  - Takabayashi, Shuji
AU  - Aoshima, Takuya
AU  - Kobayashi, Yukari
AU  - Takagi, Hisayoshi
AU  - Akasaka, Eri
AU  - Sato, Masahiro
PY  - 2020
DA  - 2020/12/03
TI  - Successful <i>i</i>-GONAD in Brown Norway Rats by Modification of <em>in vivo</em> Electroporation Conditions
JO  - OBM Genetics
SP  - 121
VL  - 04
IS  - 04
AB  - Improved-Genome editing via Oviductal Nucleic Acids Delivery (i-GONAD) was developed for in situ genome editing of the preimplantation embryos present within the oviductal lumen of mice. This method is based on intra-oviductal instillation of genome editing components and subsequent in vivo electroporation (EP) in the entire oviduct. Therefore, i-GONAD differs from the previous methods (i.e., zygote microinjection and in vitro EP) in producing genome-edited mice, which relied on ex vivo handling of preimplantation embryos and egg transfer to the recipient females. We have previously demonstrated that i-GONAD can be successfully applied to produce genome-edited rats, including albino Sprague-Dawley and albino Lewis rats (however, not pigmented Brown Norway [BN] rats). We observed that the successful i-GONAD was dependent on the mouse strain used; for example, in random-bred mice, such as ICR and C3H/He × C57BL/6, it was successful under relatively stringent electrical conditions but not in the C57BL/6 strain. Under less stringent conditions, i-GONAD was successful in the C57BL/6 strain. We speculated that this would also be true for i-GONAD using BN rats. On applying a current of >500 mA, we failed to obtain rat offspring (fetuses/newborns); however, i-GONAD under a current of 100-300 mA using NEPA21 (NEPA GENE) led to the production of genome-edited BN rats with efficiencies of 75%-100%. Similarly, i-GONAD, under a current of 150-200 mA using CUY21EDIT II (BEX Co.) led to the production of genome-edited BN rats with efficiencies of 24%-55%. These experiments suggest the importance of selecting the appropriate current value, depending on the rat strain used, when performing i-GONAD.
SN  - 2577-5790
UR  - https://doi.org/10.21926/obm.genet.2004121
DO  - 10.21926/obm.genet.2004121
ID  - Takabayashi2020
ER  -