通过玻璃化冷冻在微滴中保存的DNA注射胚胎生产转基因小鼠。

Production of transgenic mice from DNA-injected embryos cryopreserved by vitrification in microdrops.

作者信息

Landa V, Slezinger M S

机构信息

Institute of Molecular Genetics, Czechoslovak Academy of Sciences, Praha.

出版信息

Folia Biol (Praha). 1992;38(1):10-5.

PMID:1597250

Abstract

Transgenic mice were produced from DNA-injected embryos stored for 2 to 30 days in liquid nitrogen. Of the 500 zygotes collected from (C57BL/6 x CBA)F1 mice, 363 (73%) survived DNA injection into pronuclei and 246 (82%) morphologically normal 4- and 8-cell embryos were flushed from temporary recipients 48 h later. Of the 200 DNA-injected 8-cell embryos cryopreserved by vitrification in microdrops, 194 (97%) were recovered and 188 (94%) embryos were intact one hour after thawing. Of the 50 DNA-injected and frozen/thawed embryos, 48 (96%) developed to morulae or blastocysts within 30 h of in vitro culture. Transfer of 100 DNA-injected and cryopreserved 8-cell embryos into 20 day-1 recipients resulted in 47 young born. Two mice were transgenic.

摘要

转基因小鼠由在液氮中保存2至30天的注射了DNA的胚胎培育而成。从（C57BL/6×CBA）F1小鼠收集的500个受精卵中，363个（73%）在原核注射DNA后存活，48小时后从临时受体中冲出246个（82%）形态正常的4细胞和8细胞胚胎。在通过微滴玻璃化冷冻保存的200个注射了DNA的8细胞胚胎中，194个（97%）被复苏，解冻1小时后188个（94%）胚胎完整无损。在50个注射了DNA并经冷冻/解冻的胚胎中，48个（96%）在体外培养30小时内发育成桑葚胚或囊胚。将100个注射了DNA并经冷冻保存的8细胞胚胎移植到20只第1天的受体中，产下47只幼崽。其中两只小鼠为转基因小鼠。