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

CRISPR/Cas9- and siRNA-Mediated Silencing of AXL Expression.
To generate the AXL gene-edited HUVECs, an AXL-specific single-guide (sg)RNA was cloned into the lentiCRISPR v2 plasmid (Addgene; 52961). An untargeted sgRNA was used as a control. HUVECs were transduced with lentiviruses coexpressing Cas9 and sgRNA, and selected with 1.5 μg/mL puromycin at 24 h posttransduction for 4 d. To silence AXL expression, HUVECs (at 80 to 85% confluence) were transfected with 25 nM untargeted or AXL-specific siRNA using DharmaFECT 4 reagent (Dharmacon). The day after transfection, cells were detached and plated for further infection assays and the analysis of AXL cell-surface expression.

Full paper   Login or join for free to view the full paper.

Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing CRISPR Human - Deletion AXL using lentiCRISPR v2 from Addgene.

Manufacturer protocol

Download the product protocol from Addgene for lentiCRISPR v2 below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing CRISPR Human - Deletion AXL using lentiCRISPR v2 from Addgene. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.