Institute of Developmental Genetics, Helmholtz Zentrum München GmbH, German Research Center for Environmental Health, Ingolstädter Landstr. 1, D-85764 Neuherberg, Germany
Department of Molecular Hematology, University Hospital Frankfurt, Goethe University, D-60590 Frankfurt am Main, Germany.
Dis Model Mech. 2021 Feb 5;14(2):dmm047803. doi: 10.1242/dmm.047803.
Gene trapping is a high-throughput approach that has been used to introduce insertional mutations into the genome of mouse embryonic stem (ES) cells. It is performed with generic gene trap vectors that simultaneously mutate and report the expression of the endogenous gene at the site of insertion and provide a DNA sequence tag for the rapid identification of the disrupted gene. Large-scale international efforts assembled a gene trap library of 566,554 ES cell lines with single gene trap integrations distributed throughout the genome. Here, we re-investigated this unique library and identified mutations in 2202 non-coding RNA (ncRNA) genes, in addition to mutations in 12,078 distinct protein-coding genes. Moreover, we found certain types of gene trap vectors preferentially integrating into genes expressing specific long non-coding RNA (lncRNA) biotypes. Together with all other gene-trapped ES cell lines, lncRNA gene-trapped ES cell lines are readily available for functional and studies.
基因捕获是一种高通量的方法,已被用于向小鼠胚胎干细胞(ES)基因组中引入插入突变。它使用通用的基因捕获载体进行,该载体在插入位点同时突变和报告内源性基因的表达,并提供 DNA 序列标签,用于快速鉴定被破坏的基因。大规模的国际努力组装了一个包含 566554 个 ES 细胞系的基因捕获文库,其中单个基因捕获整合分布在整个基因组中。在这里,我们重新研究了这个独特的文库,除了在 12078 个独特的蛋白编码基因中发现突变外,还在 2202 个非编码 RNA(ncRNA)基因中发现了突变。此外,我们发现某些类型的基因捕获载体优先整合到表达特定长非编码 RNA(lncRNA)生物型的基因中。连同所有其他基因捕获的 ES 细胞系一起,lncRNA 基因捕获的 ES 细胞系可随时用于功能研究。