pPC1000
(Plasmid
#174829)
-
PurposeLentiviral Repair-seq vector containing CRISPRi sgRNA and SaPE2 prime edit site
-
Depositing Lab
-
Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
---|---|---|---|---|---|
Plasmid | 174829 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
-
Vector backboneLentiviral
-
Vector typeMammalian Expression, Lentiviral, CRISPR
-
Selectable markersPuromycin ; BFP
Growth in Bacteria
-
Bacterial Resistance(s)Ampicillin, 100 μg/mL
-
Growth Temperature37°C
-
Growth Strain(s)NEB Stable
-
Copy numberHigh Copy
Gene/Insert 1
-
Gene/Insert nameCRISPRi SpCas9 sgRNA cassette
-
Alt nameEGFP-targeting single guide RNA (modified F+E SpCas9 scaffold) for CRISPRi
-
SpeciesSynthetic
-
Insert Size (bp)112
-
MutationDetailed in manuscript
- Promoter mouse U6
Cloning Information for Gene/Insert 1
- Cloning method Restriction Enzyme
- 5′ cloning site BstXI (not destroyed)
- 3′ cloning site BlpI (not destroyed)
- 5′ sequencing primer mouse U6-forward (Common Sequencing Primers)
Gene/Insert 2
-
Gene/Insert namePuroR-P2A-BFP
-
Alt namePuromycin N-acetyl-transferase-P2A-blue fluorescent protein
-
SpeciesSynthetic
-
Insert Size (bp)1368
- Promoter EF1a
Cloning Information for Gene/Insert 2
- Cloning method Ligation Independent Cloning
- 5′ sequencing primer EF1a-forward
- 3′ sequencing primer WPRE-reverse (Common Sequencing Primers)
Resource Information
-
Supplemental Documents
Terms and Licenses
-
Academic/Nonprofit Terms
-
Industry Terms
- Not Available to Industry
Trademarks:
- Zeocin® is an InvivoGen trademark.
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.
-
For your Materials & Methods section:
pPC1000 was a gift from David Liu (Addgene plasmid # 174829 ; http://n2t.net/addgene:174829 ; RRID:Addgene_174829) -
For your References section:
Enhanced prime editing systems by manipulating cellular determinants of editing outcomes. Chen PJ, Hussmann JA, Yan J, Knipping F, Ravisankar P, Chen PF, Chen C, Nelson JW, Newby GA, Sahin M, Osborn MJ, Weissman JS, Adamson B, Liu DR. Cell. 2021 Oct 28;184(22):5635-5652.e29. doi: 10.1016/j.cell.2021.09.018. 10.1016/j.cell.2021.09.018 PubMed 34653350