System promises to revolutionise functional genomics and systems biology, providing a robust and simple method for site-specific disruption of DNA sequences within a host cell genome
Sigma-Aldrich announces commercialisation of the TargeTron gene knockout system.
Developed by Alan Lambowitz, University of Texas at Austin, and exclusively licensed from Ingex, the TargeTron system promises to revolutionise functional genomics and systems biology.
The TargeTron system provides the most robust and simple method available for site-specific disruption of DNA sequences within a host cell genome.
In fact, permanent gene disruption is performed in a non-random or targeted manner that does not require host recombination factors to mobilise, says Sigma-Aldrich.
The technology exploits the retrohoming ability of group II introns in order to target the exact position of gene disruption.
An extensively validated TargeTron algorithm designs specific primer sets for use in PCR mutation of the TargeTron Intron.
The mutated intron is essentially reprogrammed to insert itself into the algorithm-predicted site of any user-defined gene.
Ligation of the PCR product into a TargeTron 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.
Thus far, the technology has provided greater than 90% successfully targeted insertion rate in prokaryotes.
The current TargeTron system has been used to create permanent knockouts across a broad range of bacterial strains such as Escherichia coli, Staphylococcus aureus, Lactococcus lactis, Clostridium perfringens, Shigella flexneri and Salmonella typhimurium.
Future systems will be developed for both plant and mammalian targeted gene disruption.
"Group II intron-mediated gene disruption offers advantages over the laborious traditional gene knockout techniques such as randomly integrated transposons and cumbersome recombination methods," commented Keith Jolliff, global marketing manager of molecular biology at Sigma-Aldrich.
"TargeTron technology has potential to be the most widely applicable functional genomics tool for creating knockouts across a spectrum of host organisms."
