CRISPR-Cas9 All-in-one

The CRISPR-Cas9 All-in-One Plasmid is a highly efficient and user-friendly system for targeted gene knock-out (KO) and knock-in (KI) in mammalian cells. Originating from the prokaryotic immune system, CRISPR-Cas9 has been adapted for research applications due to its speed, cost-effectiveness, and simplicity (Nature, 2015, Vol 522, pp. 20–24). This all-in-one plasmid system features a lentiviral-compatible vector (pLenti-EF1α-Cas9-EGFP-U6-sgRNA) that co-expresses Cas9 and gene-specific guide RNA (gRNA), with EGFP included for monitoring transfection efficiency. Cas9 expression is driven by the strong EF1α promoter, while gRNA is transcribed under the U6 promoter. To use the system, researchers simply design and clone their gDNA sequence into the plasmid, transfect the target cells, visualize transfection via EGFP, and validate gene targeting using their preferred method. The system also supports delivery through AAV and lentiviral formats, and gene-specific Duo-gRNA sets are available for direct use in either cellular transfection or in vitro gRNA production. - kits