CRISPR-X helps scientists to create complex primitive genetic mutation library

Several scholars from Department of Genetics, Department of Pharmacology of Stanford University have developed a new technology of re-use of in situ protein engineering somatic hypermutation - CRISPR-X using recombinant dog proteins, which will help scientists create complex primitive genetic mutation library to analyse and improve protein engineering.

Protein engineering is currently limited by directed mutagenesis, or the construction of DNA libraries by using directed evolution to analyze protein function. This paper aims to develop a new technique for reusing somatic hypermutation for in situ protein engineering: CRISPR-X.

Researchers catalyze the activation of dormant dCas9 to induce mutations in cytidine deaminase (AID) that carry MS2-modified sgRNAs, which can specifically mutagenize endogenous targets (limiting off-target damage). This allows the generation of a number of different point mutant libraries, while targeting multiple genomic sites.

To test this approach, the researchers mutated GFP and screened for spectral shift mutations such as EGFP. And they also mutated the target of the cancer treatment drug, Bortezomib - PSMB5, which is currently the only treatment approved for malignant tumor proteasome inhibitor, which specifically inhibits chymotrypsin-like activity in the 26S proteasome of mammalian cells.

As a result, they found known and novel mutations that triggered resistance to Bortezomib. At the same time, the researchers also used the hyperactivated AID mutation to mutate the upstream and downstream sites of its transcription initiation site.

These studies show that CRISPR-X is a powerful tool that can help scientists create complex primitive mutations in the librar for analysis and improvement of protein engineering. Flarebio provides you with superior recombinant proteins like recombinant ITGB5 at competitive prices.