Disruption and sequence identification of 2,000 genes in mouse embryonic stem cells

Abstract

The dramatic increase in sequence information in the form of expressed sequence tags (ESTs) and genomic sequence has created a 'gene function gap' with the identification of new genes far outpacing the rate at which their function can be identified. The ability to create mutations in embryonic stem (ES) cells on a large scale by tagged random mutagenesis provides a powerful approach for determining gene function in a mammalian system; this approach is well established in lower organisms. Here we describe a high-throughput mutagenesis method based on gene trapping that allows the automated identification of sequence tags from the mutated genes. This method traps and mutates genes regardless of their expression status in ES cells. To facilitate the study of gene function on a large scale, we are using these techniques to create a library of ES cells called Omnibank, from which sequence-tagged mutations in 2,000 genes are described.

A stabilizing factor for mitochondrial respiratory supercomplex assembly regulates energy metabolism in muscle

“…Compared with the COX7a in complex IV, the amino-acid residues of the COX7RP protein are highly conserved 23 , suggesting its role in mitochondrial oxidative phosphorylation. We generated Cox7rpKO mice from the library of embryonic stem cells harbouring mutations by random retroviral gene trapping 24 . Genomic sequencing showed that the retroviral vector (7.4 kb) was inserted into the first intron of mouse Cox7rp, (Fig.…”

“…Cox7rpKO mice were generated by Lexicon Genetics using random retroviral gene trapping in ES cells as previously described 24 . Cox7rpKO mice were born at Mendelian ratios and were fertile.…”

Epstein‐Barr virus‐induced gene 3 negatively regulates IL‐17, IL‐22 and RORγt

“…EBI3 +/-founder mice were created at Lexicon Genetics (The Woodlands, TX) by gene trapping using random insertional mutagenesis with retroviral vector VICTR23 as described by Zambrowicz et al [31]. The integration site of the targeting cassette is located in the intronic sequence between exon 3 and exon 4 (Fig.…”

Bex1 knock out mice show altered skeletal muscle regeneration

“…OST125186, OmniBank Library, Lexicon Genetics) derived by the gene trap method [18]. Using this clone, mice heterozygous for Bex1 were generated by blastocyst injection and backcrossed to 129/SvEv mice (Taconic Farms) that were also used as controls.…”