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CRISPRPL01

Sigma-Aldrich

CRISPR GUS GAPDH Reporter Control for Monocots

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recombinant

expressed in E. coli

Quality Level

packaging

vial of 50 μL

concentration

20 ng/μL in TE buffer; DNA (1μg of plasmid DNA)

application(s)

CRISPR

selection

kanamycin

shipped in

dry ice

storage temp.

−20°C

Related Categories

General description

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

Application

  • 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.

Other Notes

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

Storage Class

12 - Non Combustible Liquids

wgk_germany

WGK 1

flash_point_f

Not applicable

flash_point_c

Not applicable


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Protocols

Zinc finger nucleases (ZFNs), were introduced by Sigma-Aldrich less than 8 years ago, but in that time the technology of targeted genome editing has advanced rapidly. Most recently, the discovery of the CRISPR/Cas9 pathway has accelerated interest in this field, opening up new possibilities for research and development.

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