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PurposeExpression of SpCas9 with 4 ERT2 fusion protein and empty gRNA cassette. The activity of Cas9 can be switched on and off in human cells with 4-hydroxytamoxifen (4-HT)
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 84232 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backboneGeneArt CRISPR Nuclease vector with OFP reporter
- Backbone size w/o insert (bp) 5081
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Vector typeMammalian Expression
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Selectable markersOFP expression
Growth in Bacteria
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Bacterial Resistance(s)Ampicillin, 100 μg/mL
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Growth Temperature37°C
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Growth Strain(s)NEB Stable
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert nameCas9
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Alt nameInducible Cas9
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SpeciesS. pyogenes
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Insert Size (bp)7969
- Promoter CMV
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Tags
/ Fusion Proteins
- ERT2-ERT2 (N terminal on insert)
- ERT2-ERT2 (C terminal on insert)
- 2A-OFP co-expression (C terminal on backbone)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site AgeI/SfoI (unknown if destroyed)
- 3′ cloning site EcoRI/XbaI (unknown if destroyed)
- 5′ sequencing primer NA (Common Sequencing Primers)
Resource Information
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Addgene Notes
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Articles Citing this Plasmid
Terms and Licenses
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Academic/Nonprofit Terms
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Industry Terms
- Not Available to Industry
Trademarks:
- Zeocin® is an InvivoGen trademark.
Depositor Comments
The four copies of ERT2 are identical in sequence and may recombine occasionally. Hence, it best to use competent cells like STBL3 or NEB Stable to propagate the plasmid and to also check the plasmid size on an agarose gel.
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
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For your Materials & Methods section:
iCas was a gift from Meng How Tan (Addgene plasmid # 84232 ; http://n2t.net/addgene:84232 ; RRID:Addgene_84232) -
For your References section:
A chemical-inducible CRISPR-Cas9 system for rapid control of genome editing. Liu KI, Ramli MN, Woo CW, Wang Y, Zhao T, Zhang X, Yim GR, Chong BY, Gowher A, Chua MZ, Jung J, Lee JH, Tan MH. Nat Chem Biol. 2016 Sep 12. doi: 10.1038/nchembio.2179. 10.1038/nchembio.2179 PubMed 27618190