Directed evolution is a powerful forward engineering approach that is heavily relied upon in protein engineering, which is essential for producing variations with enhanced performance or novel traits. Creating molecular variety at the DNA level and choosing potential proteins from a large pool of variations are the two steps in this process. However, for screening and validation, the majority of protein engineering approaches used today mostly rely on in vitro methods or microbial expression systems. The existing screening techniques may not be optimal for many protein types needing phenotypic selection in human or mammalian cells. CRISPR technology has the potential to select and diversify proteins in mammalian cells.
Introduction to CRISPR/Cas base editing enzymes
Directed evolution has already been used by nature to successfully create antibody libraries in mammalian cells. This is accomplished via somatic hypermutation, which is brought on by enzyme activation-induced cytidine deaminase (AID) in the immunoglobulin gene areas of B lymphocytes, and VDJ recombination in certain gene loci of B lymphocytes.
