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CRISPRPL01

Sigma-Aldrich

CRISPR GUS GAPDH Reporter Control for Monocots

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About This Item

UNSPSC Code:
12352200
NACRES:
NE.02

recombinant

expressed in E. coli

Quality Level

200

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

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

Legal Information

CRISPR Use License Agreement

Storage Class Code

12 - Non Combustible Liquids

WGK

WGK 1

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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Protocols

Genome Editing in Plants with CRISPR/Cas9

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.

Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.

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