Stem Cell Research and Regenerative Medicine

Gene Targeting

Gene targeting has been widely used to study human genetic diseases by removing ("knocking out"), or adding ("knocking in"), specific mutations of interest.The method are often wont to delete a gene, remove exons, add a gene and modify individual base pairs (introduce point mutations). Gene targeting can be permanent or conditional. Conditions are often a selected time during development / lifetime of the organism or limitation to a selected tissue, for instance . Gene targeting requires the creation of a selected vector for every gene of interest. However, it are often used for any gene, no matter transcriptional activity or gene size. Gene targeting methods are established for several model organisms and should vary counting on the species used. To target qualities in mice, the DNA is embedded into mouse early stage immature microorganisms in culture. Cells with the addition can add to a mouse's tissue by means of incipient organism infusion. At last, illusory mice where the altered cells structure the regenerative organs are reared. After this step the entire body of the mouse is based on the selected embryonic stem cell. To target genes in moss, the DNA is incubated along side freshly isolated protoplasts and with polyethylene glycol. As mosses are haploid organisms, moss filaments (protonema) are often directly screened for the target, either by treatment with antibiotics or with PCR. Unique among plants, this procedure for reverse genetics is as efficient as in yeast. Gene targeting has been successfully applied to cattle, sheep, swine and lots of fungi.    

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