Genome editing is a vital technology that modifies genetic information to obtain desired traits, and it is widely applied in various organisms such as microbes, plants, animals, and humans. The advent of CRISPR-Cas technology has made precise sequence editing possible by altering the target recognition sequence in guide RNA (gRNA). By expressing multiple gRNAs simultaneously, multiple targets can be edited in a single cell, significantly reducing the time and cost required to achieve specific traits.
Multiplex genome editing allows for the simultaneous editing of multiple gene sites within a single cell, greatly accelerating the process of obtaining desired traits. Precision is crucial in genome editing steps because the mismatch tolerance of the CRISPR-Cas system might cut both edited and non-edited targets, leading to cell death. Thus, using highly truncated single guide RNAs (sgRNAs) can overcome this trait of the CRISPR-Cas system. In multiplex editing, cell survival rates and editing efficiency decline as the number of targets increases.