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PurposeTo express Oct4, Klf4, Sox2, c-Myc and hairpin RNA p53. A single plasmid reprogramming system using a mini-intronic plasmid (MIP).
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 63726 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonecodon-optimized minicircle (CoMiC) construct
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Backbone manufacturerJoseph Wu's Lab at Stanford University
- Backbone size w/o insert (bp) 8062
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Modifications to backbonecodon-optimized expression cassette of the four canonical reprogramming factors OCT4, KLF4, SOX2, and c-MYC (OKSM) described previously, and combined this cassette with shRNA targeting p53
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Vector typeMammalian Expression
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Selectable markersRNA-OUT
Growth in Bacteria
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Bacterial Resistance(s)RNA-OUT ( 6% sucrose)
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Growth Temperature30°C
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Growth Strain(s)RNA-OUT selection competent cells, such as NTC4862 DH5α competent cells (Nature Technology)
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert nameMini-intronic-plasmid intron
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Alt namepUC-RNA-OUT
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Alt nameMIP intron
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SpeciesH. sapiens (human)
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Insert Size (bp)1355
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Mutationcodon-optimised
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Entrez GenePOU5F1 (a.k.a. OCT3, OCT4, OTF-3, OTF3, OTF4, Oct-3, Oct-4, Oct3/4)
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Entrez GeneKLF4 (a.k.a. EZF, GKLF)
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Entrez GeneSOX2 (a.k.a. ANOP3, MCOPS3)
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Entrez GeneMYC (a.k.a. MRTL, MYCC, bHLHe39, c-Myc)
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Entrez GeneTP53 (a.k.a. BCC7, BMFS5, LFS1, P53, TRP53)
- Promoter SFFV for Pct4, Klf4, Sox2 and c-myc; U6 for p53
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Tag
/ Fusion Protein
- IRES-dtomato (C terminal on backbone)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site AgeI (not destroyed)
- 3′ cloning site AgeI (not destroyed)
- 5′ sequencing primer unknown (Common Sequencing Primers)
Resource Information
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Supplemental Documents
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Addgene Notes
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A portion of this plasmid was derived from a plasmid made byThe pUC-RNA-OUT sequence used as MIP intron was cloned from NTC vector 8682.
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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
In addition to the publication listed above, more details can be found in Lu, et al. Mol Ther. 2013, PMID 23459514.
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:
MIP 247 CoMiP 4in1 with shRNA p53 was a gift from Mark Kay & Joseph Wu (Addgene plasmid # 63726 ; http://n2t.net/addgene:63726 ; RRID:Addgene_63726) -
For your References section:
Novel codon-optimized mini-intronic plasmid for efficient, inexpensive, and xeno-free induction of pluripotency. Diecke S, Lu J, Lee J, Termglinchan V, Kooreman NG, Burridge PW, Ebert AD, Churko JM, Sharma A, Kay MA, Wu JC. Sci Rep. 2015 Jan 28;5:8081. doi: 10.1038/srep08081. 10.1038/srep08081 PubMed 25628230