lentiCRISPR v2

CRISPR Human - Deletion AXL

Experiment
CRISPR Human - Deletion AXL
Product
lentiCRISPR v2 from Addgene
Manufacturer
Addgene

Protocol tips

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.

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Reviews

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Discussion

Discussion

1 year ago

Author: Milena Alexeyeva Russian Federation

DNA insert using CRISPR

I would like to excise a large strand of DNA and insert a new one using CRISPR. My problem is that my strand will be a little over 1kb and I am not sure if this is going to be a limiting factor. Also, how long should the homology arms be for a region of this size?

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Papers

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

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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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