pSpCas9(BB)-2A-GFP (PX458)

CRISPR Mouse - Deletion SL4 Ptgs2

Experiment
CRISPR Mouse - Deletion SL4 Ptgs2
Product
pSpCas9(BB)-2A-GFP (PX458) 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.
Ptgs2−/− SL4 cells were generated by CRISPR/Cas9-mediated genome engineering using the CRISPR design tool (www.genome-engineering.org). The genomic sequence of the Ptgs2 gene (5′-GCGGACTCCACGTGACGTAG-3′ and 5′-CAATAGTCTTAATGGCTTAC-3′) was targeted. Oligonucleotides corresponding to these guide sequences were cloned into the BbsI site of pSpCas9(BB)-2A-GFP (PX458) (Addgene), a bicistronic expression vector encoding both Cas9 and the single guide RNA. The expression vector construct was transiently transfected in SL4 cells, and then GFP-expressing cells were sorted by FACSAria (BD Biosciences).

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Reviews

pSpCas9(BB)-2A-GFP (PX458) 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 SL4 Ptgs2 using pSpCas9(BB)-2A-GFP (PX458) from Addgene.

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Manufacturer protocol

Download the product protocol from Addgene for pSpCas9(BB)-2A-GFP (PX458) below.

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Videos

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