T7-eSpCas9 plasmid

Cat: MOFY-0123-FY2

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Plasmid Information

Specifications

Species Source	Zebrafish
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Buffer	Refer to COA

Plasmid Information

Product Overview	The CRISPR/Cas genome editing tool system involves two components, sgRNA and Cas9. In the cell, the Cas9 protein recognizes and cuts the target site on the genome under the guidance of a single guide RNA (sgRNA), thereby artificially creating a double-strand break (DSB) on the genome. Once a DSB is formed, the cell's endogenous DNA damage repair mechanism will be stimulated to repair the DSB. Intracellular DNA repair includes three main forms: non-homologous end-joining repair (NHEJ), microhomology-mediated end-joining repair (MMEJ) and homologous recombination repair (HR). The first two repairs are error-prone repairs, and the result of the repair is the formation of insertion/deletion mutations near the DSB. If the insertion/deletion mutation is a frameshift mutation and occurs in the exon corresponding to the key functional domain, it may cause the target gene to be unable to encode an active protein due to the frameshift, thus becoming a null allele. It leads to gene knockout; if homologous donors are provided to the cells at the same time, these donors will have the opportunity to be used as homologous donors for DSB repair. As a result, the cells successfully achieve gene knock-in at the specified position through HR repair.
Regulatory Status	For Research Use Only
Shipping	Dry ice
Storage	Store at -20 °C.
References	1. Cong L, Ran FA, Cox D, Lin S, Barretto R, Habib N, Hsu PD, Wu X, Jiang W, Marraffini LA, Zhang F. Multiplex genome engineering using CRISPR/Cas systems. Science. 2013 Feb 15;339(6121):819-23. 2. Hsu PD, Scott DA, Weinstein JA, Ran FA, Konermann S, Agarwala V, Li Y, Fine EJ, Wu X, Shalem O, Cradick TJ, Marraffini LA, Bao G, Zhang F. DNA targeting specificity of RNA-guided Cas9 nucleases. Nat Biotechnol. 2013 Sep;31(9):827-32. 3. Adli M. The CRISPR tool kit for genome editing and beyond. Nat Commun. 2018 May 15;9(1):1911.

For Research Use Only | Not For Clinical Use.

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