lentiCRISPR v2

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	There is no need to perform a negative control golden-gate reaction (without insert) as it will always contain colonies so not a good indicator of cloning success.

Protocol tips
There is no need to perform a negative control golden-gate reaction (without insert) as it will always contain colonies so not a good indicator of cloning success.

Publication protocol

Generation of stable transfectants
CD38 was targeted for CRISPR/Cas9-mediated disruption using sgRNA sequences generated by the E-CRISP tool. Sequences were designed for use in the pLentiCRISPR v2 plasmid (Addgene #52961) (Addgene, Cambridge, MA) and obtained from IDT (Coralville, IA). Three sgRNA sequences were used (CRISPR 1, CRISPR 2, and CRISPR 3) to minimize off-target effects. The sequences are: CRISPR 1 (F: 5′-CACCGCGCCAGCAGTGGAGCGGTCC-3′; R: 5′-AAACGGACCGCTCCACTGCTGGCGC-3′), CRISPR 2 (F: 5′-CACCGCAGGGTTTGTCCCCGGACAC-3′; R: 5′-AAACGTGTCCGGGGACAAACCCTGC-3′), and CRISPR 3 (F: 5′-CACCGCTCCACTGCTGGCGCCACCT-3′; R: 5′-AAACAGGTGGCGCCAGCAGTGGAGC-3′). These sequences were each cloned into pLentiCRISPR v2 following the depositor’s protocol.

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Reviews

Discussion

Papers

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Manufacturer protocol

Download the product protocol from Addgene for lentiCRISPR v2 below.

Download manufacturer protocol

Videos

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