Why do I need to multiplex CRISPR?

CRISPR simplifies the process of targeting multiple genomic sites simultaneously. Multiplexing in CRISPR offers various benefits across several applications including: knocking out multiple genes simultaneously, carrying out genome editing with minimized off-target effects and making extensive deletions to the genome. 

• Multiplex CRISPR gene editing allows for straightforward knockout of multiple genes simultaneously by delivering multiple guide RNAs, each targeting a different gene, along with a CRISPR nuclease to the cells. 
• Additionally, multiplexing gRNAs allows for the simultaneous targeting of multiple genomic loci, enabling the modification of multiple genes within the same cell population. This can be utilized for base editing, editing CRISPRi or CRISPRa. 
• CRISPR nucleases enable the creation of extensive genomic deletions, such as gene excision and chromosomal deletion. To achieve this, at least two double-strand breaks (DSBs) flanking the targeted genomic region are required. 
• Additionally, multiplex CRISPR deletes a large region of the genome and involves precisely removing the DNA sequence situated between two specified target sites. This process typically utilizes the CRISPR-Cas9 system, where guide gRNAs are designed to guide the Cas9 nuclease to the desired locations flanking the region to be deleted.