pSpCas9(BB)-2A-Puro (PX459)

CRISPR Mouse - Deletion L929 SIRT 5

Experiment
CRISPR Mouse - Deletion L929 SIRT 5
Product
pSpCas9(BB)-2A-Puro (PX459) from Addgene
Manufacturer
Addgene

Protocol tips

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.

Publication protocol

Generation of CRISPR/Cas9-mediated knockout cell lines
L929 cells were transfected with pSpCas9-(BB)2A puro (PX459), a plasmid from Feng Zhang's lab,68 and knockout cells were selected using puromycin at 10 μg/ml for 5 days. Guide oligos used were mouse Sirt2 (5′ primer: CAC CGG CGG AAG TCA GGG ATA CCC G; 3′ primer: AAA CCG GGT ATC CCT GAC TTC CGC C), and mouse Sirt5 (5′ primer: CAC CGA ACT GGG AAA TGA ATC GGC C; 3′ primer: AAA CGG CCG ATT CAT TTC CCA GTT C).



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Reviews

pSpCas9(BB)-2A-Puro (PX459) from Addgene has not yet been reviewed for this experiment

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Discussion

Discussion

4 years ago

Author: Mario Udinese Italy

Floxing mice with CRISPR

Hi everyone! I am planning on floxing mice with CRISPR but I am having trouble deciding which region to target. Do you have any tips on choosing?

Discussion

5 years ago

Author: Ben Saar Israel

How to choose a region to target for CRISPR

Hi everyone! I am planning on floxing mice with CRISPR but I am having trouble deciding which region to target. Do you have any tips on choosing?

Share your thoughts or question with experts in your field by adding a discussion!

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing CRISPR Mouse - Deletion L929 SIRT 5 using pSpCas9(BB)-2A-Puro (PX459) from Addgene.

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Manufacturer protocol

Download the product protocol from Addgene for pSpCas9(BB)-2A-Puro (PX459) below.

Download PDF Download manufacturer protocol

Videos

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