The all in one vector (Lenti-EF1a-Cas9-EGFP-U6 sgRNA) from ARP is designed for the expression of Cas9 and gRNA for mammalian gene modification studies using CRISPR/Cas9 technology in vivo or in vitro.
Cas9 is expressed from a bidirectional EF1a promoter and sgRNA is cloned downstream of a human U6 promote. To construct the vector, you must construct pGL3-U6 sgRNA-Puro with your guide sequence. The U6-gRNA sequence is obtained by PCR, and the PCR production is cloned into the pre-linearized vector using the included high-efficiency ligation mix and competent cells. After injected with the vector, sgRNA can recognize the targeted DNA sequence and guide Cas9 nuclease during genome editing for gene knockout, knockin, mutagenesis, and more.