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CAS9MRNA

Sigma-Aldrich

Cas9 mRNA

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packaging

vial of 50 μL

concentration

500 ng/μL

shipped in

dry ice

storage temp.

−70°C

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This Item
CAS9D10APRCAS9PCAS9BST
vibrant-m

CAS9MRNA

Cas9 mRNA

vibrant-m

CAS9D10APR

Cas9-D10A Nickase Protein

vibrant-m

CAS9P

Cas9 plasmid

vibrant-m

CAS9BST

CAS9 Blasticidin Lenti Plasmid

storage temp.

−70°C

storage temp.

−20°C

storage temp.

−20°C

storage temp.

−20°C

concentration

500 ng/μL

concentration

-

concentration

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

concentration

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

shipped in

dry ice

shipped in

dry ice

shipped in

dry ice

shipped in

dry ice

packaging

vial of 50 μL

packaging

pkg of 1 kit (3 components)

packaging

vial of 50 μL

packaging

vial of 50 μL

General description

Cas9 mRNA generated through in-vitro transcription which has been capped and polyA-tailed. This mRNA should be used in conjunction with purified guideRNA. The use of RNA vs. plasmid constructs avoids complications with promoter-embryo compatibility as well as the possibility of random integration of nuclease and gRNA-expressing plasmids into the host animal genome.

Application

Functional Genomics/Transgenic Applications
  • Creation of gene knockouts in transgenic applications
  • Creation of knock-in animals with promoters, fusion tags or reporters integrated into endogenous genes

Features and Benefits

RNA ready to use in transgenic applications. Must be used in conjunction with a purified guideRNA sequence to mediate a site specific double strand break in the DNA.

Components

1 vial containing 25ug of Cas9 mRNA.

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

Principle

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.

Physical form

Sigma Cas9 mRNA is supplied at a concentration of 500 ng/ul (50 ul Cas9 mRNA, capped and polyA-tailed)

Preparation Note

While Sigma implements the same high quality RNA synthesis procedures that have been successful in many mRNA-based ZFN transgenic projects, we highly recommend centrifuging your final, concentration-adjusted CRISPR RNA samples immediately prior to microinjection to ensure all particulate material has been removed which may result in clogging of microinjection needles.

Other Notes

Must be used in conjunction with the RNA format of a guideRNA sequence in order to mediate a double strand break in the DNA.
Typical microinjection concentrations used in the literature are in the ranges of 20-200 ng/ul for Cas9 mRNA and 10-50 ng/ul for gRNA.
While Sigma CRISPR RNA was not developed for robust application to cell culture, we have seen some success in detecting CEL-I activity using RNA-only formats for both CRISPR nucleases and paired nickases.  RNA-only delivery formats may be favorable for cell types which are sensitive to double stranded DNA such a dendritic cells or when promoter-cell incompatibilities exist.

Storage Class

12 - Non Combustible Liquids

wgk_germany

WGK 1

flash_point_f

Not applicable

flash_point_c

Not applicable


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Marc-Olivier Turgeon et al.
Endocrinology, 158(4), 815-830 (2017-03-23)
Loss-of-function mutations in the X-linked immunoglobulin superfamily, member 1 (IGSF1) gene cause central hypothyroidism. IGSF1 is a transmembrane glycoprotein of unknown function expressed in thyrotropin (TSH)-producing thyrotrope cells of the anterior pituitary gland. The protein is cotranslationally cleaved, with only

Protocols

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.

Related Content

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).

Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.

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