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CRISPR/Cas9 Ribonucleoprotein-mediated Precise Gene Editing by Tube Electroporation
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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.
Cryo Letters
|October 26, 2023
Summary
Vitrification of bovine zygotes (early-stage embryos) before or after genome editing via electroporation did not impact editing efficiency. However, vitrification did reduce overall embryonic development rates.
Area of Science:
- Reproductive biology
- Genetics
- Embryology
Background:
- Cryopreservation of bovine zygotes enables flexible genome editing schedules.
- Vitrification may cause cell membrane damage, potentially affecting embryonic development and genome mutation.
Purpose of the Study:
- To assess the impact of vitrification timing on bovine zygote development and genome mutation after electroporation.
- To compare genome editing outcomes when vitrification precedes or follows electroporation.
Main Methods:
- In vitro-derived bovine zygotes were subjected to CRISPR/Cas9 genome editing.
- Electroporation was performed either immediately after vitrification/warming (Vitrified-EP), 2 hours after (Vitrified-2h-EP), or before vitrification (EP-vitrified).
Main Results:
- Vitrified-warmed zygotes showed significantly lower development rates compared to controls.
- No significant differences in mutation rates or efficiency were observed in blastocysts, regardless of electroporation timing relative to vitrification.
Conclusions:
- Vitrification timing (before or after electroporation) does not influence the efficiency of genome editing in bovine zygotes.
- While genome editing is unaffected, vitrification negatively impacts overall zygote development.
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