Development and Genome Mutation of Bovine Zygotes Vitrified Before and After Genome Editing via Electroporation

IF 1 4区生物学 Q3 BIOLOGY

Cryo letters Pub Date : 2023-03-01 DOI:10.54680/fr23210110612

T. Nguyen, L. Do, Z. Namula, Qingyi Lin, N. Torigoe, M. Nagahara, M. Hirata, F. Tanihara, T. Otoi

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引用次数: 0

Abstract

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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电穿孔基因组编辑前后玻璃化牛受精卵的发育和基因组突变

背景:牛受精卵的低温保存允许通过电穿孔进行基因组编辑的灵活时间表。然而，玻璃化诱导的细胞膜损伤不仅会影响胚胎发育，还会导致基因组突变。目的:探讨电穿孔前后受精卵玻璃化处理对牛推定受精卵发育和基因组突变的影响。材料和方法:体外衍生的牛受精卵用CRISPR/Cas9系统立即(玻璃化- ep)或孵育后2小时(玻璃化-2h- ep)在玻璃化和加热后电穿孔，或在玻璃化前电穿孔(ep -玻璃化)。结果:玻璃化加热受精卵发育率显著低于未玻璃化受精卵发育率(p < 0.05)。此外，无论电穿孔处理的时间如何，电穿孔受精卵产生的囊胚的突变率和突变效率都没有差异。结论:我们的研究结果表明，在电穿孔治疗前后玻璃化不影响受精卵的基因组编辑。

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来源期刊

Cryo letters 生物-生理学

CiteScore

1.80

自引率

10.00%

发文量

审稿时长

1 months

期刊介绍： A bimonthly international journal for low temperature sciences, including cryobiology, cryopreservation or vitrification of cells and tissues, chemical and physical aspects of freezing and drying, and studies involving ecology of cold environments, and cold adaptation The journal publishes original research reports, authoritative reviews, technical developments and commissioned book reviews of studies of the effects produced by low temperatures on a wide variety of scientific and technical processes, or those involving low temperature techniques in the investigation of physical, chemical, biological and ecological problems.