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CRISPR01 Sigma-Aldrich

CRISPR Human EMX1 Positive Control

  •  NACRES NA.51

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Properties

Related Categories CRISPR Controls (DNA and Virus), CRISPR-Cas9, Functional Genomics and RNAi, Molecular Biology
Quality Level   200
packaging   pkg of 2 vials (50μL aliquot for each of the 2 kit components)
concentration   20 ng/μL in TE buffer; DNA (1μg of plasmid DNA)
application(s)   CRISPR: suitable
shipped in   dry ice
storage temp.   −20°C

Description

General description

Validated CRISPR site, which serves as an experimental control for the Wt Cas9. A two component positive control system consisting of a CMV-driven Cas9 plasmid and a U6-driven guide RNA plasmid targeting the human EMX1 gene.

Application

Functional Genomics/Target Validation
• Creation of gene knockouts in cell lines
• Creation of knock-in cell lines with promoters, fusion tags or reporters integrated into endogenous genes

Features and Benefits

Serves as an experimental control for the CRISPR editing workflow using WT Cas9. Allows for validation of your system with the CRISPR/Cas9 system. A positive result in a miss-match detection assay will indicate validation of your system.

Components

1 vial containing 1ug of U6-gRNA plasmid expressing a guide sequence to human EMX1. 1 vial containing 1ug of Cas9 plasmid.

Principle

CRISPR/Cas systems are employed by bacteria and archaea as a defense against invading viruses and plasmids. Recently, the type II CRISPR/Cas system from the bacterium Streptococcus pyogenes has been engineered to function in eukaryotic systems using two molecular components: a single Cas9 protein and a non-coding guide RNA (gRNA). The Cas9 endonuclease can be programmed with a single gRNA, directing a DNA double-strand break (DSB) at a desired genomic location. Similar to DSBs induced by zinc finger nucleases (ZFNs), the cell then activates endogenous DNA repair processes, either non-homologous end joining (NHEJ) or homology-directed repair (HDR), to heal the targeted DSB.

Physical form

Sigma U6-gRNA plasmid expressing a guide sequence to human EMX1 supplied at a concentration of 20ng/ul in 50ul. Sigma Cas9 plasmid at a concentration of 20ng/ul in 50ul.

Preparation Note

Sigma CRISPR plasmid products are delivered as mini-prep aliquots,
which may not be suitable for transfection into particular cell types. For best results, we advise maxi-prepping
plasmids using endotoxin-free DNA purification kits prior to transfection.

Other Notes

Typical transfection concentrations used in literature are in the ranges of >= 1.0ug/uL and <= 5uL of Cas9 plasmid combined with >= 1.0ug/uL and <= 5uL of U6-gRNA plasmids. (All dosages above assume 0.5 to 1 million cells nucleofected)

Legal Information

CRISPR Use License Agreement

Lentiviral and WPRE License Agreements

Safety & Documentation

Safety Information

RIDADR 
NONH for all modes of transport

Documents

Certificate of Analysis (COA)

Please Enter a Lot Number
Protocols & Articles

Articles

A CRISPR/Cas-GFP Vector for Rapid Expression Verification and Enrichment of Genome Edited Cells

In many genome editing experiments involving ZFNs and CRISPR/Cas nucleases, the first challenge is achieving successful delivery of plasmids and subsequent expression of the encoded nucleases. While ...
Keywords: Cloning, Gene expression, Sequencing, Transfection

CRISPR/Cas Nuclease RNA-guided Genome Editing

What is CRISPR/Cas9? How does CRISPR/Cas work? How does CRISPR Cas9 work? What is CRISPR/Cas9? Our role in developing CRISPR/Cas9
Keywords: Acetylations, Catalysis, Cell culture, Degradations, Gene expression, Genetic, PAGE, Polymorphisms, Transcription, Transduction, Transfection

Tips for Cell Engineering using Cas9-GFP CRISPR plasmids

CRISPR endonucleases have shown wide variation in their activity, even among multiple CRISPRs designed within close genomic proximity.1  For this reason, we highly recommend that you test 3 to 4 CRIS...
Keywords: Cell culture, Cloning, DNA purification, Gene expression, Microscopy, Purification, Reductions, Sequencing, Transcription, Transfection

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CRISPR

In recent years CRISPR has revolutionized gene editing capabilities, leading to sophisticated ways to create success with any experiment. As the first company to offer custom biomolecules globally fo...
Keywords: Genetic, Genomics, Transduction, Transfection

Predictive Models for Neuroscience using CRISPR [VIDEO]

Caroline Beckett, the global CRISPR product manager, discusses reagent solutions for creating predictive models for neuroscience research. She notes that many neuroscientists want to be able to creat...
Keywords: Cell culture, Diseases, Neurodegenerative Diseases, Neuroscience

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