CRISPR GUS GAPDH Reporter Control for Dicots CRISPRPL02-1EA

NACRES:

NA.51

All-in-one, ready-to-use Cas9 and guide RNA (gRNA) expression plasmids with GUS Reporter
CRISPR 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

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

Features and Benefits

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

Components
1 vial containing 50ul of 20ng/ul plasmid DNA
Keep reagent tubes closed when not in use.
Practice aseptic lab technique to avoid DNase contamination.
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.
CRISPR Use License Agreement,

Quality Level	200,
recombinant	expressed in E. coli
packaging	vial of 50 µL
concentration	20 ng/µL in TE buffer; DNA (1µg of plasmid DNA)
application(s)	CRISPR: suitable
selection	kanamycin
shipped in	dry ice
storage temp.	20°C

RIDADR	NONH for all modes of transport
WGK Germany	WGK 1
Flash Point F	Not applicable
Flash Point C	Not applicable