通过电穿孔在基因组编辑前后进行玻璃化。

IF 1 4区 生物学 Q3 BIOLOGY Cryo letters Pub Date : 2023-03-01
T V Nguyen, L T Kim Do, Z Namula, Q Y Lin, N Torigoe, M Nagahara, M Hirata, F Tanihara, T Bio-Innovation Research Center Tokushima University Tokushima Japan Otoi
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引用次数: 0

摘要

背景:牛受精卵的冷冻保存允许通过电穿孔进行灵活的基因组编辑。然而,玻璃化诱导的细胞膜损伤不仅可能影响胚胎发育,还可能影响基因组突变。目的:探讨电穿孔处理前后受精卵玻璃化对牛受精卵发育和基因组突变的影响。材料和方法:体外来源的牛受精卵在玻璃化和加温后立即用CRISPR/Cas9系统(玻璃化EP)或孵育2小时后(玻璃化-2h-EP)电穿孔,结果:玻璃化温合子的发育率明显低于未玻璃化的对照合子(p<0.05)。此外,无论电穿孔处理的时间如何,电穿孔合子产生的胚泡的突变率和突变效率都没有观察到差异。结论:我们的研究结果表明,电穿孔处理前后的玻璃化处理不影响合子的基因组编辑。
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Vitrified before and after genome editing via electroporation.

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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来源期刊
Cryo letters
Cryo letters 生物-生理学
CiteScore
1.80
自引率
10.00%
发文量
50
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.
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