pX330-U6-Chimeric_BB-CBh-hSpCas9

CRISPR Mouse - Activation C2C12 Lama1

Experiment
CRISPR Mouse - Activation C2C12 Lama1
Product
pX330-U6-Chimeric_BB-CBh-hSpCas9 from Addgene
Manufacturer
Addgene

Protocol tips

Protocol tips
The presence of PEG and glycerol (supplied by the Ligation Buffer and
the T4 DNA Ligase) will make the reaction mixture viscous. Be sure to mix the reaction thoroughly by pipetting up and down. Do not vortex.

Publication protocol

Expression Vector
The plasmid px330-U6-Chimeric-HsPCas9 (Addgene plasmid #42230) was used for expression of the gRNA and of dCas9. It contains two BbsI restriction sites for insertion of an oligonucleotide, which selects the target sequence of the gRNA. This gRNA is under the control of the U6 promoter. The plasmid pAC154-dual-dCas9-VP160-sgExpression (Addgene plasmid #48240) was used for expression of the gRNA and dCas9-VP160. It contains two BbsI restriction sites for insertion of an oligonucleotide, which selects the target sequence of the gRNA. This gRNA is under the control of the U6 promoter. This plasmid was modified to include a gene encoding resistance to puromycin, the latter is separated from the gene encoding dCas9 via a sequence coding for the 2A peptide. There is a nuclear localization sequence (NLS) fused with the dCas9-VP160 gene.

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Reviews

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Discussion

Discussion

1 year 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

2 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 - Activation C2C12 Lama1 using pX330-U6-Chimeric_BB-CBh-hSpCas9 from Addgene.

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Manufacturer protocol

Download the product protocol from Addgene for pX330-U6-Chimeric_BB-CBh-hSpCas9 below.

Download PDF Download manufacturer protocol

Videos

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