PEA1-Nuclease-Puro
(Plasmid
#171992)
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PurposeDelivers all prime editing nuclease components in a single, puromycin selectable plasmid
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 171992 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepCMV-PE2
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Vector typeMammalian Expression
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Selectable markersPuromycin
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 nameCbH-Cas9-RT-T2A-Puro, hU6-pegRNA (Bbs1 gate), hU6-sgRNA (Bbs1 gate)
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SpeciesSynthetic; Cas9 is from S. pyogenes; M-MLV RT is from the Moloney murine leukemia virus
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MutationGC to CA point mutation in SpCas9 to restore nuclease function
- Promoter CMV for Cas9, U6 for gRNAs
Cloning Information
- Cloning method Ligation Independent Cloning
- 5′ sequencing primer --
- 3′ sequencing primer BGH-reverse (Common Sequencing Primers)
Resource Information
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Supplemental Documents
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
Full sequence https://benchling.com/s/seq-zQAEVlUC4JpjV6Gt4RFT.
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:
PEA1-Nuclease-Puro was a gift from Paul Thomas (Addgene plasmid # 171992 ; http://n2t.net/addgene:171992 ; RRID:Addgene_171992) -
For your References section:
Optimized nickase- and nuclease-based prime editing in human and mouse cells. Adikusuma F, Lushington C, Arudkumar J, Godahewa GI, Chey YCJ, Gierus L, Piltz S, Geiger A, Jain Y, Reti D, Wilson LOW, Bauer DC, Thomas PQ. Nucleic Acids Res. 2021 Sep 17. pii: 6371974. doi: 10.1093/nar/gkab792. 10.1093/nar/gkab792 PubMed 34534334