lentiCRISPR v2

CRISPR Human - Deletion TLN2

Experiment

CRISPR Human - Deletion TLN2

Product

lentiCRISPR v2 from Addgene

Manufacturer

Addgene

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Tips Publication protocol Reviews Discussion Papers Manufacturer protocol Videos

Protocol tips

Upstream tips	Protocol tips	Downstream tips
To access a plasmid, keep the plate on dry ice to prevent thawing. Using a sterile pipette tip (20uL or 200uL one), puncture the seal above an individual well and spread a portion of the glycerol stock onto an agar plate.	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.

Upstream tips
To access a plasmid, keep the plate on dry ice to prevent thawing. Using a sterile pipette tip (20uL or 200uL one), puncture the seal above an individual well and spread a portion of the glycerol stock onto an agar plate.
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

Knockout of TLN1 and TLN2 with CRISPR
Preparation of viruses and cell infection were performed as described previously (Li et al., 2013; Wu et al., 2011). LentiCRISPR-TLN1 and lentiCRISPR-TLN2 were co-transfected with packaging vectors pMDLg/pRRE, pRSV-Rev and CMV-VSVG into 293FT cells. Lentiviral particles were collected and used to infect U2OS cells. The cells were selected with puromycin, and clones were isolated. TLN1- or TLN2-knockout clones were detected by western blotting using anti-talin1 and anti-talin2 monoclonal antibodies.

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Reviews

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Discussion

4 years 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 TLN2 using lentiCRISPR v2 from Addgene.

Paper title

Talin2-mediated traction force drives matrix degradation and cell invasion.

Full paper

Manufacturer protocol

Download the product protocol from Addgene for lentiCRISPR v2 below.

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Videos

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