CAS9GFPP
CMV-CAS9-2A-GFP Plasmid
Synonyme(s) :
Cas9 plasmid
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About This Item
Code UNSPSC :
12352200
Nomenclature NACRES :
NA.51
Produits recommandés
Produit recombinant
expressed in E. coli
Niveau de qualité
Conditionnement
vial of 50 μL
Concentration
20 ng/μL in TE buffer; DNA (1μg of plasmid DNA)
Promoteur
Promoter name: CMV
Gène reporter
GFP
selection
kanamycin
Conditions d'expédition
dry ice
Température de stockage
−20°C
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Catégories apparentées
Description générale
The Cas9 expression plasmids use the CMV promoter for strong transient expression of Cas9. Alternate promoters can be substituted by replacement of CMV using MluI and NheI. Also, the Cas9 expression plasmids can be linearized using XbaI for T7-based mRNA production. The addition of a fluorophore that is translationally co-expressed with the Cas9 nuclease allows for easy visualization of successful transfection.
Application
CMV-CAS9-2A-GFP plasmid has been used to induce additional sex combs-like1 mutations in human U937 leukemic cells. It has also been used in CRISPR/Cas9 analysis.
CMV-CAS9-2A-GFP plasmid is suitable for functional genomics/target validation for:
CMV-CAS9-2A-GFP plasmid is suitable for functional genomics/target validation for:
- Creation of gene knockouts in multiple cell lines
- Complete knockout of genes not amenable to RNAi
- Creation of knock-in cell lines with promoters, fusion tags, or reporters integrated into endogenous genes
Composants
1 vial containing 1 μg of Cas9-2A-GFP plasmid.
Please note, product does not contain guideRNA sequence. This must be purchased separately through the Custom CRISPR product tab.
Please note, product does not contain guideRNA sequence. This must be purchased separately through the Custom CRISPR product tab.
Principe
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.
Autres remarques
Must be used in conjunction with a U6-gRNA plasmid in order to mediate a double strand break in the DNA.
Typical transfection concentrations used in literature are in the ranges of >= 1.0 μg/μL and <= 5 μL of Cas9 plasmid combined with >= 1.0 μg/μL and <= 5 μL of U6-gRNA plasmids. (All dosages above assume 0.5 to 1 million cells nucleofected).
Typical transfection concentrations used in literature are in the ranges of >= 1.0 μg/μL and <= 5 μL of Cas9 plasmid combined with >= 1.0 μg/μL and <= 5 μL of U6-gRNA plasmids. (All dosages above assume 0.5 to 1 million cells nucleofected).
Informations légales
Code de la classe de stockage
10 - Combustible liquids
Classe de danger pour l'eau (WGK)
nwg
Point d'éclair (°F)
Not applicable
Point d'éclair (°C)
Not applicable
Certificats d'analyse (COA)
Recherchez un Certificats d'analyse (COA) en saisissant le numéro de lot du produit. Les numéros de lot figurent sur l'étiquette du produit après les mots "Lot" ou "Batch".
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Targeting synthetic lethality between the SRC kinase and the EPHB6 receptor may benefit cancer treatment.
Paul J M, et al.
Oncotarget, 7(31), 50027-50027 (2016)
CRISPR/Cas9-mediated ASXL1 mutations in U937 cells disrupt myeloid differentiation.
Wu Z J, et al.
International Journal of Oncology, 52(4), 1209-1223 (2018)
A Scotto Rosato et al.
Nature communications, 10(1), 5630-5630 (2019-12-12)
The lysosomal calcium channel TRPML1, whose mutations cause the lysosomal storage disorder (LSD) mucolipidosis type IV (MLIV), contributes to upregulate autophagic genes by inducing the nuclear translocation of the transcription factor EB (TFEB). Here we show that TRPML1 activation also
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