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 STING-deficient cells with CRISPRs.
A single guide RNA (sgRNA) targeting STING exon 5 (sgRNASTING) (5′-AATATGACCATGCCAGCCCA-3′) was cloned into the LentiCRISPR vector (catalog number 49535; Addgene) (26) to generate pLVCR-sgSTING. Human telomerase reverse transcriptase (hTERT)-immortalized HUVECs (tHUVECs) were transduced with either the empty LentiCRISPR vector (control) or pLVCR-sgSTING (STING knockout [STINGKO]) and selected with puromycin (1 μg/ml) for 7 days. To obtain single-cell clones, cells were plated at 0.6 cells per well into 96-well plates, and all STINGKO clones used in this study (clones 9 and 11) were verified by Sanger sequencing for deletion and by Western blotting for STING expression.

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Papers

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Manufacturer protocol

Download the product protocol from Addgene for lentiCRISPR v2 below.

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Videos

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