JDS246

CRISPR Mouse - Deletion C2C12 Prnp

Experiment
CRISPR Mouse - Deletion C2C12 Prnp
Product
JDS246 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 gRNA Expression Vectors
The SpCas9 plasmid JDS246 (Plasmid 43861) and the gRNA expression plasmid MLM3636 (Plasmid 43860) were obtained from a non-profit plasmid share repository (Addgene, Cambridge, MA, USA). Suitable CRISPR target sites within Exon 3 positive and negative strands were identified using the ‘CRISPR Design Tool’ () described in the Results section. The respective oligonucleotide pairs were obtained from Life Technologies (Burlington, ON, Canada) and were customized to include overhangs compatible for ligation into MLM3636 linearized by digestion with BsmB1 (cat. no. R0580S; New England BioLabs, Ipswich, MA, USA), a cloning site located in this vector on the 3′ side of a U6 promoter element. Oligonucleotides were phosphorylated with polynucleotide kinase (cat. no. EK0031; Fermentas, Ottawa, ON, Canada), annealed and inserted into the gRNA plasmid using T4 DNA ligase (cat no. M0202S; New England Biolabs) and transformed into Turbo competent (cat no. C2984H; New England Biolabs).

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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 - Deletion C2C12 Prnp using JDS246 from Addgene.

Paper title
CRISPR-Cas9-Based Knockout of the Prion Protein and Its Effect on the Proteome
Full paper
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Manufacturer protocol

Download the product protocol from Addgene for JDS246 below.

Download PDF Download manufacturer protocol

Videos

Check out videos that might be relevant for performing CRISPR Mouse - Deletion C2C12 Prnp using JDS246 from Addgene. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.

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