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The CRISPR-Cas method allows genes contained within a cell to be modified. As a general rule, the method could previously only be used to modify a single gene. Basel-based researchers have now successfully refined the method, allowing them to apply it to 25 – or more – genes at the same time.
The CRISPR-Cas method has been used in the scientific world for several years now to precisely and “comparatively easily” remove, replace or modify genes contained within a cell, as outlined in a press release issued by the Swiss Federal Institute of Technology in Zurich (ETH). Up until now, however, the method has, generally speaking, only allowed a single cell to be modified. On occasion, researchers successfully managed to modify two or three in one go, although ETH also explained in the press release that researchers were able to modify seven genes simultaneously in one very particular case. However, researchers working at the Department of Biosystems Science and Engineering (D-BSSE) at ETH, which is based in Basel, have now revolutionized this method which has already proven so successful.
The team headed up by Randall Platt has now developed a process that allows the CRISPR-Cas method to modify “25 target sites within genes in a cell at once”. According to Platt, the new method even has the potential to be upscaled to “dozens or even hundreds of genes”. The D-BSSE professor commented in the press release: “Thanks to this new tool, we and other scientists can now achieve what we could only dream of doing in the past”.
The new method exploits the fact that genes connect and form networks with one another. “Our method enables us, for the first time, to systematically modify entire gene networks in a single step”, Platt explains. The increase or reduction in activity of certain genes facilitated by the CRISPR-Cas method can now be applied by the Basel researchers to complete networks.
To this end, the researchers have created a plasmid, or a circular DNA molecule, that not only stores the blueprint of the Cas enzyme, but also contains a sort of “longer address list”, allowing several genes to be modified. Furthermore, for the new technique, the scientists did not use the Cas9 enzyme that has featured in most CRISPR-Cas methods to date, but rather the Cas12a enzyme. In the press release, ETH explains how this enzyme is involved in the method by allowing “address labels” to be cut from the address list. “The shorter these addressing sequences are, the more of them we can fit onto a plasmid”, Platt says.