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CAS9RFPP

Sigma-Aldrich

CMV-CAS9-2A-RFP Plasmid

Sinónimos:

Cas9 plasmid

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About This Item

Código UNSPSC:
41106610
NACRES:
NA.51

recombinante

expressed in E. coli

Nivel de calidad

200

envase

vial of 50 μL

concentración

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

Promotor

Promoter name: CMV

gen reportero

RFP

Condiciones de envío

dry ice

temp. de almacenamiento

−20°C

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Descripción general

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.

Aplicación

Functional Genomics/Target Validation
  • 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

Componentes

1 vial containing 1 μg of Cas9-2A-RFP plasmid.

Please note, product does not contain guideRNA sequence. This must be purchased separately through the Custom CRISPR product tab.

Principio

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.

Forma física

Sigma Cas9-2A-RFP plasmid DNA is supplied at concentrations of 50ng/μl in 20μl.

Otras notas

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).

Información legal

CRISPR Use License Agreement

Código de clase de almacenamiento

10 - Combustible liquids

Clase de riesgo para el agua (WGK)

nwg

Punto de inflamabilidad (°F)

Not applicable

Punto de inflamabilidad (°C)

Not applicable

Certificados de análisis (COA)

Busque Certificados de análisis (COA) introduciendo el número de lote del producto. Los números de lote se encuentran en la etiqueta del producto después de las palabras «Lot» o «Batch»

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Artículos

Tips for Cell Engineering Using Cas9-GFP CRISPR Plasmids

CRISPR endonucleases have shown wide variation in their activity, even among multiple CRISPRs designed within close genomic proximity.

CRISPR/Cas-GFP Vectors for Rapid Expression Verification and Enrichment of Genome Edited Cells

View experimental data showing crispr/cas expression and enrichment using FACS

Protocolos

CRISPR Cas 9 Nuclease RNA-guided Genome Editing

Learn about CRISPR Cas9, what it is and how it works. CRISPR is a new, affordable genome editing tool enabling access to genome editing for all.

Contenido relacionado

Gene Expression & Silencing

Sigma-Aldrich® Advanced Genomics is the leading provider of gene editing and silencing technologies including CRISPR, Cas9, synthetic guide RNA (sgRNA), and Zinc Finger Nuclease (ZFN).

Nuestro equipo de científicos tiene experiencia en todas las áreas de investigación: Ciencias de la vida, Ciencia de los materiales, Síntesis química, Cromatografía, Analítica y muchas otras.

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