Application

To verify successful integration of T-DNA in plant genome GUS receptor wheat gGAPDH control for monocots for Agrobacterium mediated transformation

Components

1 vial containing 50ul of 20ng/ul plasmid DNAKeep reagent tubes closed when not in use. Practice aseptic lab technique to avoid DNase contamination.

Features and Benefits

Low cost, genome editing option compared to other methods. Easy to use Online ordering Ready to ship in 2 days

General description

All-in-one, ready-to-use Cas9 and guide RNA (gRNA) expression plasmids with GUS ReporterCRISPR Plant Cas9 products are intended for Agrobacterium-mediated plant transformation. The products are based on the type IIA CRISPR-Cas9 derived from Streptococcus pyogenes. The native Cas9 coding sequence is codon optimized for expression in monocots and dicots, respectively. The monocot Cas9 constructs contain a monocot U6 promoter for sgRNA expression, and the dicot Cas9 constructs contain a dicot U6 promoter. Arabidopsis seedlings were germinated in 6 well tissue culture plates. The seedlings were infected with Agrobacterium which had the CRISPR plasmids with a GUS reporter. After 3-4 days of transfection the GUS expression was detected. b-glucuronidase (GUS) is an enzyme that hydrolyzes colorless glucuronides to yield colored product

Legal Information

CRISPR Use License Agreement

Other Notes

For ordering any of our custom CRISPR plant products please visit: CUSTOM ORDERING FORM

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.