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Updated: Jul 12, 2025
CRISPR/Cas9 Ribonucleoprotein-mediated Precise Gene Editing by Tube Electroporation
Published on: June 20, 2019
Vitrified before and after genome editing via electroporation
T V Nguyen1, L T Kim Do2, Z Namula3
1Faculty of Veterinary Medicine, Vietnam National University of Agriculture, 100000 Hanoi, Vietnam; Bio-Innovation Research Center, Tokushima University, 7793233 Tokushima, Japan.
Background:
Cryopreservation of bovine zygotes allows for a flexible schedule of genome editing via electroporation. However, vitrification-induced cell membrane damage may not only affect embryonic development but also genome mutation.
Objective:
To investigate the effects of vitrification of zygotes before and after electroporation treatments on the development and genome mutation of bovine presumptive zygotes.
Materials And Methods:
In vitro-derived bovine zygotes were electroporated with the CRISPR/Cas9 system immediately (Vitrified-EP) or 2 h after incubation (Vitrified-2h-EP) following vitrification and warming, or electroporated before vitrification (EP-vitrified).
Results:
The development rates of vitrified-warmed zygotes were significantly lower (p < 0.05) than those of control zygotes that were not vitrified. Moreover, no differences were observed in the mutation rates and mutation efficiency of the blastocysts resulting from electroporated zygotes, irrespective of the timing of electroporation treatment.
Conclusion:
Our results suggest that vitrification before and after electroporation treatments does not affect the genome editing of zygotes.
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