Biology: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 et al. (2005). "Tagging genes with cassette-exchange sites". Nucleic Acids Res 33 (4): e44. doi:10.1093/nar/gni045. PMID 15741177.
- ↑ De-Zolt, S 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. PMID 16478711.
- ↑ "IGTC, International Gene Trap Consortium". http://www.genetrap.org/.
Further reading
- "Mouse embryonic stem cells and reporter constructs to detect developmentally regulated genes". Science 244 (4903): 463–5. April 1989. doi:10.1126/science.2497519. PMID 2497519.
- "Identification of cellular promoters by using a retrovirus promoter trap". Journal of Virology 63 (8): 3227–33. August 1989. doi:10.1128/JVI.63.8.3227-3233.1989. PMID 2545900.
- "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. doi:10.1101/gad.5.9.1513. PMID 1653172.
- "Disruption and sequence identification of 2,000 genes in mouse embryonic stem cells". Nature 392 (6676): 608–11. April 1998. doi:10.1038/33423. PMID 9560157.
- "Establishment of a gene-trap sequence tag library to generate mutant mice from embryonic stem cells". Nature Genetics 24 (1): 13–4. January 2000. doi:10.1038/71622. PMID 10615117.
- "Gene-trap mutagenesis: past, present and beyond". Nature Reviews Genetics 2 (10): 756–68. October 2001. doi:10.1038/35093548. PMID 11584292.
- "A public gene trap resource for mouse functional genomics". Nature Genetics 36 (6): 543–4. June 2004. doi:10.1038/ng0604-543. PMID 15167922.
- "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. doi:10.1073/pnas.1633296100. PMID 12904583.
- "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. doi:10.1073/pnas.2336103100. PMID 14610273.
- "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. doi:10.1073/pnas.0502273102. PMID 15870191.
- "Gene traps: tools for plant development and genomics". The Plant Cell 12 (7): 1007–20. July 2000. doi:10.2307/3871251. PMID 10899970.
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