Gene trapping
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]
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
- 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. 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.
- 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.
- 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.
- 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.