Gene trapping explained

Gene trapping is a high-throughput approach that is used to introduce insertional mutations across an organism's genome.

Method

Trapping is performed with gene trap vectors whose principal element is a gene trapping cassette consisting of a promoterless reporter gene and/or selectable genetic marker, flanked by an upstream 3' splice site (splice acceptor; SA) and a downstream transcriptional termination sequence (polyadenylation sequence; polyA).

When inserted into an intron of an expressed gene, the gene trap cassette is transcribed from the endogenous promoter of that gene in the form of a fusion transcript in which the exon(s) upstream of the insertion site is spliced in frame to the reporter/selectable marker gene. Since transcription is terminated prematurely at the inserted polyadenylation site, the processed fusion transcript encodes a truncated and nonfunctional version of the cellular protein and the reporter/selectable marker. Thus, gene traps simultaneously inactivate and report the expression of the trapped gene at the insertion site, and provide a DNA tag (gene trap sequence tag, GTST) for the rapid identification of the disrupted gene.^{[1] [2]}

Access

The International Gene Trap Consortium is centralizing the data and supplies modified cell lines.^[3]

Further reading

• Gossler A, Joyner AL, Rossant J, Skarnes WC . Mouse embryonic stem cells and reporter constructs to detect developmentally regulated genes . Science . 244 . 4903 . 463–5 . April 1989 . 2497519 . 10.1126/science.2497519.
• von Melchner H, Ruley HE . Identification of cellular promoters by using a retrovirus promoter trap . Journal of Virology . 63 . 8 . 3227–33 . August 1989 . 10.1128/JVI.63.8.3227-3233.1989 . 2545900 . 250892 .
• Friedrich G, Soriano P . Promoter traps in embryonic stem cells: a genetic screen to identify and mutate developmental genes in mice . Genes & Development . 5 . 9 . 1513–23 . September 1991 . 1653172 . 10.1101/gad.5.9.1513. free .
• Zambrowicz BP, Friedrich GA, Buxton EC, Lilleberg SL, Person C, Sands AT . Disruption and sequence identification of 2,000 genes in mouse embryonic stem cells . Nature . 392 . 6676 . 608–11 . April 1998 . 10.1038/33423 . 9560157 . 4329895 .
• Wiles MV, Vauti F, Otte J, Füchtbauer EM, Ruiz P, Füchtbauer A, Arnold HH, Lehrach H, Metz T, von Melchner H, Wurst W . Establishment of a gene-trap sequence tag library to generate mutant mice from embryonic stem cells . Nature Genetics . 24 . 1 . 13–4 . January 2000 . 10.1038/71622 . 10615117 . 29505338 .
• Stanford WL, Cohn JB, Cordes SP . Gene-trap mutagenesis: past, present and beyond . Nature Reviews Genetics . 2 . 10 . 756–68 . October 2001 . 10.1038/35093548 . 11584292 . 11468334 .
• Skarnes WC, von Melchner H, Wurst W, Hicks G, Nord AS, Cox T, Young SG, Ruiz P, Soriano P, Tessier-Lavigne M, Conklin BR, Stanford WL, Rossant J . A public gene trap resource for mouse functional genomics . Nature Genetics . 36 . 6 . 543–4 . June 2004 . 2716026 . 10.1038/ng0604-543 . 15167922 .
• Hansen J, Floss T, Van Sloun P, Füchtbauer EM, Vauti F, Arnold HH, Schnütgen F, Wurst W, von Melchner H, Ruiz P . A large-scale, gene-driven mutagenesis approach for the functional analysis of the mouse genome . Proceedings of the National Academy of Sciences of the United States of America . 100 . 17 . 9918–22 . August 2003 . 10.1073/pnas.1633296100 . 12904583 . 187885 . free .
• Zambrowicz BP, Abuin A, Ramirez-Solis R, Richter LJ, Piggott J, BeltrandelRio H, Buxton EC, Edwards J, Finch RA, Friddle CJ, Gupta A, Hansen G, Hu Y, Huang W, Jaing C, Key BW, Kipp P, Kohlhauff B, Ma ZQ, Markesich D, Payne R, Potter DG, Qian N, Shaw J, Schrick J, Shi ZZ, Sparks MJ, Van Sligtenhorst I, Vogel P, Walke W, Xu N, Zhu Q, Person C, Sands AT . 6 . Wnk1 kinase deficiency lowers blood pressure in mice: a gene-trap screen to identify potential targets for therapeutic intervention . Proceedings of the National Academy of Sciences of the United States of America . 100 . 24 . 14109–14 . November 2003 . 10.1073/pnas.2336103100 . 14610273 . 283554 . free .
• Schnütgen F, De-Zolt S, Van Sloun P, Hollatz M, Floss T, Hansen J, Altschmied J, Seisenberger C, Ghyselinck NB, Ruiz P, Chambon P, Wurst W, von Melchner H . Genomewide production of multipurpose alleles for the functional analysis of the mouse genome . Proceedings of the National Academy of Sciences of the United States of America . 102 . 20 . 7221–6 . May 2005 . 1129123 . 10.1073/pnas.0502273102 . 15870191 . free .
• Springer PS . Gene traps: tools for plant development and genomics . The Plant Cell . 12 . 7 . 1007–20 . July 2000 . 10899970 . 149045 . 10.2307/3871251. 3871251 .

Notes and References

1. G. Cobellis. G. Nicolaus. etal. 2005. Tagging genes with cassette-exchange sites. Nucleic Acids Res. 33. 4. e44. 15741177. 10.1093/nar/gni045. 552971.
2. S. De-Zolt. F. Schnutgen. etal. 2006. High-throughput trapping of secretory pathway genes in mouse embryonic stem cells. Nucleic Acids Res. 34. 3. 25. 16478711. 10.1093/nar/gnj026. 1369290.
3. Web site: IGTC, International Gene Trap Consortium. www.genetrap.org. 2018-02-11.