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
{"title":"通过电穿孔在基因组编辑前后进行玻璃化。","authors":"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","doi":"","DOIUrl":null,"url":null,"abstract":"<p><strong>Background: </strong>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.</p><p><strong>Objective: </strong>To investigate the effects of vitrification of zygotes before and after electroporation treatments on the development and genome mutation of bovine presumptive zygotes.</p><p><strong>Materials and methods: </strong>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).</p><p><strong>Results: </strong>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.</p><p><strong>Conclusion: </strong>Our results suggest that vitrification before and after electroporation treatments does not affect the genome editing of zygotes.</p>","PeriodicalId":10937,"journal":{"name":"Cryo letters","volume":null,"pages":null},"PeriodicalIF":1.0000,"publicationDate":"2023-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Vitrified before and after genome editing via electroporation.\",\"authors\":\"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\",\"doi\":\"\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Background: </strong>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.</p><p><strong>Objective: </strong>To investigate the effects of vitrification of zygotes before and after electroporation treatments on the development and genome mutation of bovine presumptive zygotes.</p><p><strong>Materials and methods: </strong>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).</p><p><strong>Results: </strong>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.</p><p><strong>Conclusion: </strong>Our results suggest that vitrification before and after electroporation treatments does not affect the genome editing of zygotes.</p>\",\"PeriodicalId\":10937,\"journal\":{\"name\":\"Cryo letters\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":1.0000,\"publicationDate\":\"2023-03-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Cryo letters\",\"FirstCategoryId\":\"99\",\"ListUrlMain\":\"\",\"RegionNum\":4,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q3\",\"JCRName\":\"BIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Cryo letters","FirstCategoryId":"99","ListUrlMain":"","RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"BIOLOGY","Score":null,"Total":0}
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.
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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