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PurposeExpresses FRB fused SpCas9 for NanoMEDIC packaging.
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Depositing Lab
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Depositing OrganizationKyoto University, Center for iPS Cell Research and Application
Located in Japan -
Citations
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Sequence Information
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 138477 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
| DNA | 138477-D50 | 50 μg of DNA in Tris buffer | $495 | ||
| DNA | 138477-D100 | 100 μg of DNA in Tris buffer | $585 | ||
Backbone
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Vector backbonepHLS
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Backbone manufacturerHotta lab
- Backbone size w/o insert (bp) 4762
- Total vector size (bp) 9007
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Modifications to backbonepHL vector with SV40 replication origin.
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Vector typeMammalian Expression, CRISPR
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)DH5alpha
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert nameSpCas9, human codon optimized
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Alt nameCRISPR-Cas9
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SpeciesStreptococcus pyogenes
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Tags
/ Fusion Proteins
- FRB (N terminal on insert)
- 3x HA tag (C terminal on insert)
Resource Information
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.
DNA (Catalog # 138477-D50)
Purpose
Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.
Delivery
- Amount 50 μg
- Concentration 1 μg/μL
- Pricing $495 USD
- Storage 4 ℃ (short-term) or -20 ℃ (long-term)
Terms and Licenses
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Academic/Nonprofit Terms
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Industry Terms
- Not Available to Industry
Quality Control
Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.
Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.
DNA (Catalog # 138477-D100)
Purpose
Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.
Delivery
- Amount 100 μg
- Concentration 1 μg/μL
- Pricing $585 USD
- Storage 4 ℃ (short-term) or -20 ℃ (long-term)
Terms and Licenses
-
Academic/Nonprofit Terms
-
Industry Terms
- Not Available to Industry
Quality Control
Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.
Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.
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:
pHLS-EF1a-FRB-SpCas9-A was a gift from Akitsu Hotta (Addgene plasmid # 138477 ; http://n2t.net/addgene:138477 ; RRID:Addgene_138477) -
For your References section:
Extracellular nanovesicles for packaging of CRISPR-Cas9 protein and sgRNA to induce therapeutic exon skipping. Gee P, Lung MSY, Okuzaki Y, Sasakawa N, Iguchi T, Makita Y, Hozumi H, Miura Y, Yang LF, Iwasaki M, Wang XH, Waller MA, Shirai N, Abe YO, Fujita Y, Watanabe K, Kagita A, Iwabuchi KA, Yasuda M, Xu H, Noda T, Komano J, Sakurai H, Inukai N, Hotta A. Nat Commun. 2020 Mar 13;11(1):1334. doi: 10.1038/s41467-020-14957-y. 10.1038/s41467-020-14957-y PubMed 32170079