SMS2 CRISPR/Cas9 KO Plasmid (m)

CRISPR Mouse - Deletion C2C12 Sgms2

Experiment
CRISPR Mouse - Deletion C2C12 Sgms2
Product
SMS2 CRISPR/Cas9 KO Plasmid (m) from Santa Cruz Biotechnology
Manufacturer
Santa Cruz Biotechnology

Protocol tips

Upstream tips
If transfecting more than one plasmid (i.e., CRISPR/Cas9 KO Plasmid with HDR Plasmid), mix Plasmid DNA at equivalent ratios.
Protocol tips
Do not add antibiotics to the Plasmid Transfection Medium

Publication protocol

Generation of SMS1 and SMS2 KO Cells Using CRISPR/Cas9 KO System
Sphingomyelin synthase 1 (Sgms1) and 2 (Sgms2) CRISPR/Cas9 knock-out plasmids (sc-431565 and sc-428888) were purchased from Santa Cruz Biotechnology, Inc. We followed the manufacturer's protocol. Briefly, C2C12 myoblasts were seeded in a 6-well plate in 3 ml of antibiotic-free 10% FBS DMEM, a day prior to transfection. When cells reached ∼50% confluence, the plasmid DNA solution composed of 2 μg of CRISPR/Cas9 KO plasmid and 2 μg of the corresponding HDR plasmid (sc-431565-HDR and sc-428888-HDR) in 15 μl of UltraCruz transfection reagent (sc395739) was added dropwise to each well. The cells were incubated and monitored for 24–72 h. Successful co-transfection of the CRISPR/Cas9 KO plasmid and HDR plasmid was confirmed by detection of red fluorescent protein under a fluorescence microscope, and the cells were selected with media containing puromycin to generate the KO cell line. After 1 week of incubation with puromycin, the gene expression was assessed by RT-qPCR following RNA isolation and cDNA synthesis. Primer sequences used herein correspond to the gRNA sequences in the CRISPR/Cas9 KO plasmids.

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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 Sgms2 using SMS2 CRISPR/Cas9 KO Plasmid (m) from Santa Cruz Biotechnology.

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Manufacturer protocol

Download the product protocol from Santa Cruz Biotechnology for SMS2 CRISPR/Cas9 KO Plasmid (m) below.

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Videos

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