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CRISPR06

Sigma-Aldrich

CRISPR UNIVERSAL NEGATIVE CONTROL 1

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NACRES:
NA.51

form

liquid

packaging

vial of 50 μL

concentration

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

application(s)

CRISPR

shipped in

dry ice

storage temp.

−20°C

Related Categories

General description

Universal negative control CRISPRs have been designed not to recognize any sequence in the human, mouse or rat genome. A single vector format is provided which includes the Cas9 expression cassette and gRNA sequence. This vector includes GFP which is co-expressed from the same mRNA as the Cas9 protein via a 2A peptide linkage and enables for tracking of transfection efficiency or enrichment in cell populations via FACS.

Application

Functional Genomics/Target Validation
  • Creation of gene knockouts in cell lines
  • Creation of knock-in cell lines with promoters, fusion tags or reporters integrated into endogenous genes

Features and Benefits

Allows for expression of Cas9 and GFP without specific targeting of the CRISPR/Cas9 system.

Components

1 vial containing 1ug of U6-gRNA/CMV-Cas9-GFP plasmid expressing a non-targeting guide sequence.

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 U6-gRNA/CMV-Cas9-GFP plasmid expressing a non-targeting guide sequence supplied at a concentration of 20ng/ul in 50ul.

Preparation Note

Sigma CRISPR plasmid products are delivered as mini-prep aliquots,
which may not be suitable for transfection into particular cell types. For best results, we advise maxi-prepping
plasmids using endotoxin-free DNA purification kits prior to transfection.

Other Notes

Typical transfection concentrations used in literature are in the ranges of 2.0 to 8.0ug of the single vector expressing the guideRNA and Cas9 . (All dosages above assume 0.5 to 1 million cells nucleofected)

Legal Information

CRISPR Use License Agreement

Lentiviral and WPRE License Agreements

Storage Class Code

12 - Non Combustible Liquids

WGK

WGK 1

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Regulatory Information

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Molecular cancer research : MCR, 17(2), 508-520 (2018-09-30)
Isocitrate dehydrogenase 1 (IDH1) is the most commonly mutated metabolic enzyme in human malignancy. A heterozygous genetic alteration, arginine 132, promotes the conversion of α-ketoglutarate to D-2-hydroxyglutarate (2-HG). Although pharmacologic inhibitors of mutant IDH1 are promising, resistance mechanisms to targeted
Takahiro Yamazaki et al.
Nature immunology, 21(10), 1160-1171 (2020-08-05)
Autophagy supports both cellular and organismal homeostasis. However, whether autophagy should be inhibited or activated for cancer therapy remains unclear. Deletion of essential autophagy genes increased the sensitivity of mouse mammary carcinoma cells to radiation therapy in vitro and in
Nathalie Falk et al.
Journal of cell science, 131(16) (2018-07-29)
Pericentrin (Pcnt) is a multifunctional scaffold protein and mutations in the human PCNT gene are associated with several diseases, including ciliopathies. Pcnt plays a crucial role in ciliary development in olfactory receptor neurons, but its function in the photoreceptor-connecting cilium
Aitziber Buqué et al.
Nature communications, 11(1), 3819-3819 (2020-08-01)
Hormone receptor (HR)+ breast cancer (BC) causes most BC-related deaths, calling for improved therapeutic approaches. Despite expectations, immune checkpoint blockers (ICBs) are poorly active in patients with HR+ BC, in part reflecting the lack of preclinical models that recapitulate disease

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Protocols

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