Germline Transmission of a Novel Rat Embryonic Stem Cell Line Derived from Transgenic Rats
Germline Transmission of a Novel Rat Embryonic Stem Cell Line Derived from Transgenic Rats
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DOI：
10.1089/scd.2012.0040
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发表时间：
2012-09-01
影响因子：
4
通讯作者：
Bryda, Elizabeth C.
中科院分区：
文献类型：
--
作者：
Men, Hongsheng;Bauer, Beth A.;Bryda, Elizabeth C.
种系胜任型大鼠胚胎干（ES）细胞系是利用ES细胞基因靶向技术建立突变大鼠模型的重要资源。从任何大鼠品系（包括转基因品系）中分离出具有生殖系能力的胚胎干细胞细胞系的能力，将允许进行更复杂的基因操作，而无需大量繁殖。以携带增强型绿色荧光蛋白（EGFP）转基因的雄性斯普拉格·道利（SD）作为胚胎干细胞系衍生的创始动物。建立了许多胚胎干细胞系，并进行了严格的质量控制测试，包括多能因子表达评估、核型分析和病原体/无菌测试。两个雄性胚胎干细胞系，SD-Tg。EC1/Rrrc和SD-Tg。EC8/Rrrc被注射到雄性与雌性杂交的黑鼠（DA）囊胚中。将嵌合动物与野生型SD交配，通过鉴定携带胚胎干细胞衍生的EGFP转基因的幼崽，验证胚胎干细胞系的种系传递性。虽然两种胚胎干细胞系都能产生嵌合动物，但只有SD-Tg。EC1具有种系能力。这证实了从转基因大鼠菌株中获得生殖系胜任的胚胎干细胞系的可行性，并为未来的遗传操作创造新的大鼠模型提供了一种具有稳定绿色荧光蛋白（GFP）报告基因的新型胚胎干细胞系。
Germline-competent rat embryonic stem (ES) cell lines are important resources for the creation of mutant rat models using ES-cell-based gene targeting technology. The ability to isolate germline-competent ES cell lines from any rat strain, including genetically modified strains, would allow for more sophisticated genetic manipulations without extensive breeding. Sprague Dawley (SD) males carrying an enhanced green fluorescent protein (EGFP) transgene were used as the founder animals for the derivation of ES cell lines. A number of ES cell lines were established and subjected to rigorous quality control testing that included assessment of pluripotency factor expression, karyotype analysis, and pathogen/sterility testing. Two male ES cell lines, SD-Tg.EC1/Rrrc and SD-Tg.EC8/Rrrc, were injected into blastocysts recovered from a cross of Dark Agouti (DA) males with SD females. Resulting chimeric animals were bred with wild-type SD mates to verify the germline transmissibility of the ES cell lines by identifying pups carrying the ES cell line-derived EGFP transgene. While both ES cell lines gave rise to chimeric animals, only SD-Tg.EC1 was germline competent. This confirms the feasibility of deriving germline-competent ES cell lines from transgenic rat strains and provides a novel ES cell line with a stable green fluorescent protein (GFP) reporter for future genetic manipulations to create new rat models.