CRISPR Human EMX1 Positive Control

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

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.

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.

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.

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

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)

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.

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.