Cas9-D10A nickase - P&A Biotech

Our Price	$199
Quantity	50μg (1mg/mlx50μl, 313pmol)
Catalog number	P15002
Description	Cas9(CRISPR associated protein 9) mutant (D10A), csn1, SPy_1046 Cas9 Nickase is a Cas9 mutant(D10A). 2 gRNA spacers are needed. It has better specificity for gene knock-in through a HDR donor, which has 2 guides for the cleavage site. Cas9 and cas9 nickase both have similar efficiency to cleave the target, but cas9 nickase has a lower off-target effect.
Application	Cas9 nickase can be used for 1) In vitro DNA cleavage by gRNA guiding; 2) Genome editing through the injection for mammalian zygote, zebrafish embryos, & drosophila embryos; 3) Introduction into cell by transfection. In this buffer, Cas9 can directly be used for microinjection, eletroporation and in vitro DNA cleavage
Source	Humanized Cas9 nickase gene from an E. Coli K-12 strain.
Physical appearance	In buffer[20 mM Hepes pH 7.5, 150 mM KCl, 1 mM MgCl2, 5%glycerol, 1 mM Tris(2-chloroethyl) phosphate (TCEP)].
Formulation	The protein concentrations can be at 5mg/ml (equal to 31μM), or at 1mg/ml (equal to 6.3μM) in a micro-injection buffer.
Solubility	The Cas9 nickase is very soluble in water and most aqueous buffers.
Stability	Cas9 nickase is stable at room temperature, but should be stored desiccated below 0°C.
Purity	Greater than 98.0% as determined by SDS-PAGE.
Amino acid sequence	The N terminus contains 6xHis-tag and the C-terminus contains TAT peptide for the nuclear localization signal(NLS) and cell-penetrating peptide(CPP).
Biological activity	An assay of biological activity is done by in vitro gRNA guide DNA cleavage.
Protein content	Protein purity of 98%. Quantification was carried out by Bradford methods.
References	1. ``Efficient Intracellular Delivery of Native Protein``. Cell, April 23, 2015 ( deliver Cas9 protein and gRNA for stem cell) 2.``Generation of knock-in primary human T cells using Cas9 ribonucleoproteins``. The Doudna lab and Marson lab, PNAS. June 29, 2015 3. ``Gene disruption by cell-penetrating peptide-mediated delivery of Cas9 protein and guide RNA`` Genome Research. 2014
Usage	P & A Biotech’s products are furnished for laboratory research use only.