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.12
Access
The International Gene Trap Consortium is centralizing the data and supplies modified cell lines.3
References
References
- Cobellis, G; Nicolaus, G; et al. (2005). "Tagging genes with cassette-exchange sites". Nucleic Acids Res. 33 (4): e44. doi:10.1093/nar/gni045. PMC 552971. PMID 15741177.
- De-Zolt, S; Schnutgen, F; et al. (2006). "High-throughput trapping of secretory pathway genes in mouse embryonic stem cells". Nucleic Acids Res. 34 (3): 25. doi:10.1093/nar/gnj026. PMC 1369290. PMID 16478711.
- "IGTC, International Gene Trap Consortium". www.genetrap.org. Retrieved 2018-02-11.
Further reading
Further reading
- Gossler A, Joyner AL, Rossant J, Skarnes WC (April 1989). "Mouse embryonic stem cells and reporter constructs to detect developmentally regulated genes". Science. 244 (4903): 463–5. doi:10.1126/science.2497519. PMID 2497519.
- von Melchner H, Ruley HE (August 1989). "Identification of cellular promoters by using a retrovirus promoter trap". Journal of Virology. 63 (8): 3227–33. doi:10.1128/JVI.63.8.3227-3233.1989. PMC 250892. PMID 2545900.
- Friedrich G, Soriano P (September 1991). "Promoter traps in embryonic stem cells: a genetic screen to identify and mutate developmental genes in mice". Genes & Development. 5 (9): 1513–23. doi:10.1101/gad.5.9.1513. PMID 1653172.
- Zambrowicz BP, Friedrich GA, Buxton EC, Lilleberg SL, Person C, Sands AT (April 1998). "Disruption and sequence identification of 2,000 genes in mouse embryonic stem cells". Nature. 392 (6676): 608–11. doi:10.1038/33423. PMID 9560157. S2CID 4329895.
- Wiles MV, Vauti F, Otte J, Füchtbauer EM, Ruiz P, Füchtbauer A, Arnold HH, Lehrach H, Metz T, von Melchner H, Wurst W (January 2000). "Establishment of a gene-trap sequence tag library to generate mutant mice from embryonic stem cells". Nature Genetics. 24 (1): 13–4. doi:10.1038/71622. PMID 10615117. S2CID 29505338.
- Stanford WL, Cohn JB, Cordes SP (October 2001). "Gene-trap mutagenesis: past, present and beyond". Nature Reviews Genetics. 2 (10): 756–68. doi:10.1038/35093548. PMID 11584292. S2CID 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 (June 2004). "A public gene trap resource for mouse functional genomics". Nature Genetics. 36 (6): 543–4. doi:10.1038/ng0604-543. PMC 2716026. PMID 15167922.
- Hansen J, Floss T, Van Sloun P, Füchtbauer EM, Vauti F, Arnold HH, Schnütgen F, Wurst W, von Melchner H, Ruiz P (August 2003). "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. doi:10.1073/pnas.1633296100. PMC 187885. PMID 12904583.
- Zambrowicz BP, Abuin A, Ramirez-Solis R, Richter LJ, Piggott J, BeltrandelRio H, et al. (November 2003). "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. doi:10.1073/pnas.2336103100. PMC 283554. PMID 14610273.
- 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 (May 2005). "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. doi:10.1073/pnas.0502273102. PMC 1129123. PMID 15870191.
- Springer PS (July 2000). "Gene traps: tools for plant development and genomics". The Plant Cell. 12 (7): 1007–20. doi:10.2307/3871251. JSTOR 3871251. PMC 149045. PMID 10899970.