CRISPR GUS GAPDH Reporter Control for Monocots

200

expressed in E. coli

vial of 50 μL

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

CRISPR

kanamycin

dry ice

−20℃

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

1管含50μl的20ng/μl质粒DNA
在不使用时，将试管盖扣紧。
利用无菌实验技术，避免DNAase污染。

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.

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