Gene targeting
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Gene targeting is a genetic technique that uses homologous recombination to change an endogenous gene. The method can be used to delete a gene, remove exons, and introduce point mutations. Gene targeting can be permanent or conditional. Conditions can be a specific time during development / life of the organism or limitation to a specific tissue, for example. Gene targeting requires the creation of a specific vector for each gene of interest. However, it can be used for any gene, regardless of transcriptional activity or gene size.
Method
Gene targeting methods vary depending on organism. To target genes in mice a rough outline of the necessary steps is as follows: First, a targeting construct made out of DNA is generated in bacteria. It typically contains part of the gene to be targeted, a reporter gene, and a selectable marker. This construct is then inserted into mouse embryonic stem cells in culture. After cells with the correct insertion have been selected, they can be used to contribute to a mouse's tissue via embryo injection. Finally, chimeric mice where the modified cells made up the reproductive organs are selected for via breeding. After this step the entire body of the mouse is based on the previously selected embryonic stem cell.
Comparison with gene trapping
Gene trapping is based on random insertion of a 80's cassette while gene targeting targets a specific gene. 80's cassettes can be used for many different things while the flanking homology regions of gene targeting cassettes need to be adapted for each gene. This makes gene trapping more easily amenable for large scale projects than targeting. On the other hand, gene targeting can be used for genes with low transcriptions that would go undetected in a trap screen. Also, the probability of trapping increases with intron size. For gene targeting these compact genes are just as easily altered.
2007 Nobel prize
Mario R. Capecchi, Martin J. Evans and Oliver Smithies were declared laureates of the 2007 Nobel Prize in Physiology or Medicine for their work on "principles for introducing specific gene modifications in mice by the use of embryonic stem cells", or gene targeting.[1]
References
See also
- genetics, genetic engineering
- gene knockout, knockout mouse
- gene trapping (random gene knockout technique)
- gene knockdown (reduction not removal of a gene product)
- Mus musculus (house mouse; common model organism)
- Toll-like receptor (example of a gene targeted for analysis)
External links
- guide to gene targeting by the University of California, San Diego
- outline of gene targeting by the University of Michigan
- gene targeting diagram & summary by Heydari lab, Wayne State Universityid:Penargetan gen
it:Gene targeting sl:Ciljanje genov Jan!!!
Acknowledgement and Attribution Regarding Sources of Content
Some of the initial content on this page may be incorporated in part from copyleft sources in the public domain including wikis such as Wikipedia and AskDrWiki. Drug information for patients came from the The National Library of Medicine. Infectious disease information may have come from the Centers for Disease Control (CDC). Differential Diagnoses are drawn from clinicians as well as an amalgamation of 3 sources: 1.The Disease Database; 2. Kahan, Scott, Smith, Ellen G. In A Page: Signs and Symptoms. Malden, Massachusetts: Blackwell Publishing, 2004:3; 3. Sailer, Christian, Wasner, Susanne. Differential Diagnosis Pocket. Hermosa Beach, CA: Borm Bruckmeir Publishing LLC, 2002:7 .
