CRISPR07

UNIWERSALNA KONTROLA NEGATYWNA CRISPR 2

• Kod UNSPSC: 41106609
• NACRES: NA.51

Właściwości

• Formularz: liquid
• Poziom jakości: 200
• opakowanie: vial of 50 μL
• stężenie: 20 ng/μL in TE buffer; DNA (1μg of plasmid DNA)
• Zastosowanie: CRISPR
• Warunki transportu: dry ice
• temp. przechowywania: −20°C

Opis

Opis ogólny

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.

Zastosowanie

Genomika funkcjonalna/walidacja celu

• Tworzenie nokautów genów w liniach komórkowych
• Tworzenie linii komórkowych knock-in z promotorami, znacznikami fuzyjnymi lub reporterami zintegrowanymi z endogennymi genami

Cechy i korzyści

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

Komponenty

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

Zasada

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.

Postać fizyczna

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

Uwaga dotycząca przygotowania

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.

Inne uwagi

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)

Informacje prawne

CRISPR Use License Agreement

Lentiviral and WPRE License Agreements

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Informacja o środkach bezpieczeństwa

• Kod klasy składowania: 12 - Non Combustible Liquids
• Klasa zagrożenia wodnego (WGK): WGK 1
• Temperatura zapłonu (°F): Not applicable
• Temperatura zapłonu (°C): Not applicable