Gene trapping


Gene trapping

Gene trapping is a high-throughput approach that is used to introduce insertional mutations across the mammalian genome. It 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 non-functional 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.An international public consortium [http://www.genetrap.org/ International Gene Trap Consortium] is centralizing the data and cell lines can be requested from them

References

article to read:
# Gossler, A., A. L. Joyner, et al. (1989). "Mouse embryonic stem cells and reporter constructs to detect developmentally regulated genes." Science 244(4903): 463-5.
# von Melchner, H. and H. E. Ruley (1989). "Identification of cellular promoters by using a retrovirus promoter trap." J Virol 63(8): 3227-33.
# Zambrowicz, B. P., G. A. Friedrich, et al. (1998). "Disruption and sequence identification of 2,000 genes in mouse embryonic stem cells." Nature 392(6676): 608-11.
# Wiles, M. V., F. Vauti, et al. (2000). "Establishment of a gene-trap sequence tag library to generate mutant mice from embryonic stem cells." Nat Genet 24(1): 13-4.
# Gene Trap Mutagenesis past, present & beyond; Stanford WL, Cohn JB, Cordes SP (2001)Nature Rev. Genet., 2, 756–768
# Skarnes, W. C., H. von Melchner, et al. (2004). "A public gene trap resource for mouse functional genomics." Nat Genet 36(6): 543-4.
# Hansen, J., T. Floss, et al. (2003). "A large-scale, gene-driven mutagenesis approach for the functional analysis of the mouse genome." Proc Natl Acad Sci U S A 100(17): 9918-22.
# Zambrowicz, B. P., A. Abuin, et al. (2003). "Wnk1 kinase deficiency lowers blood pressure in mice: A gene-trap screen to identify potential targets for therapeutic intervention." Proc Natl Acad Sci U S A 100(24): 14109-14.
# Schnutgen, F., S. De-Zolt, et al. (2005). "Genomewide production of multipurpose alleles for the functional analysis of the mouse genome." Proc Natl Acad Sci U S A 102(20): 7221-
# De-Zolt, S., F. Schnutgen, et al. (2006). "High-throughput trapping of secretory pathway genes in mouse embryonic stem cells." Nucleic Acids Res 34(3): e25.
# Patricia S. Springer (2000). "Gene Traps: Tools for Plant Development and Genomics." The Plant Cell, Vol. 12:1007-1020.


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