The TargeTron gene-knockout system provides a simple method for site-specific disruption of DNA sequences within a host cell genome. Permanent gene disruption is performed in a nonrandom or targeted manner that does not require host recombination factors to mobilize. The technology exploits the retrohoming ability of group II introns to target the exact position of gene disruption. A validated TargeTron algorithm designs specific primer sets for use in PCR mutation of the TargeTron Intron. The mutated intron is reprogrammed to insert itself into the algorithm-predicted site of any user-defined gene. Ligation of the PCR product into a Targe Tron vector is followed by transformation of the host cell. Expression results in a ribonucleoprotein complex that targets the intron to the precise insertion site within the host genome, resulting in gene knockout.
Sigma-Aldrich
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