Abstract

Somatic Cell Nuclear Transfer (SCNT) technique was used to produce the first viable cloned cattle offspring in Russia. Whole-genome SNP genotyping confirmed that the cloned calf was identical to the fibroblast cell line that was used for SCNT. CRISPR/Cas9 approach was subsequently used to knock out genes for beta-lactoglobulin gene (PAEP) and the beta-lactoglobulin-like protein gene (LOC100848610) in the fibroblast cells. Gene editing (GE) efficiency was 4.4% for each of these genes. We successfully obtained single-cell-derived fibroblast colonies containing PAEP and LOC100848610 knockouts, which will be used to produce beta-lactoglobulin-deficient cattle.

中文翻译：

---

牛胚胎成纤维细胞的克隆后代生产和CRISPR / Cas9基因组编辑

摘要

体细胞核移植（SCNT）技术被用于生产俄罗斯第一个可行的克隆牛后代。全基因组SNP基因分型证实，克隆的小牛与用于SCNT的成纤维细胞系相同。随后使用CRISPR / Cas9方法敲除成纤维细胞中β-乳球蛋白基因（PAEP）和β-乳球蛋白样蛋白基因（LOC100848610）的基因。这些基因每个的基因编辑（GE）效率是4.4％。我们成功地获得了包含PAEP和LOC100848610基因敲除物的单细胞衍生成纤维细胞菌落，这些菌落将用于生产β-乳球蛋白缺陷型牛。