PurposeExpresses dead Cas9 (dCas9) fused to DNMT3A catalytic domain (CD) linked to IRES-mCherry under the control of TetOp and a minimal CMV promoter
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||78254||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
Vector backboneAddgene # 20321
- Total vector size (bp) 14829
Modifications to backboneWe took out OSKM and put in new MCS site: EcoRI-XbaI-NheI-AgeI-PspXI-AscI-EcoRI
Vector typeMammalian Expression, CRISPR
Growth in Bacteria
Bacterial Resistance(s)Ampicillin, 100 μg/mL
Copy numberHigh Copy
Gene/Insert nameDNMT3A CD aa 598-912
SpeciesH. sapiens (human)
Insert Size (bp)948
Entrez GeneDNMT3A (a.k.a. DNMT3A2, HESJAS, M.HsaIIIA, TBRS)
- Promoter CMV
/ Fusion Protein
- Flag epitope tag (N terminal on insert)
- Cloning method Restriction Enzyme
- 5′ cloning site NheI (not destroyed)
- 3′ cloning site AgeI (not destroyed)
- 5′ sequencing primer ctgcagccttcaagtacttcgacacc
- 3′ sequencing primer ATGCTCGTCAAGAAGACAGGGCCAGGTTTC (Common Sequencing Primers)
Article Citing this Plasmid
Terms and Licenses
- Not Available to Industry
- Zeocin® is an InvivoGen trademark.
The dCas9 insert in this plasmid also contains a T1167A mutation, which is unlikely to affect the function of the plasmid, as it is already a catalytically dead Cas9 mutant.
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:TetO-dCas9-D3A was a gift from Grant Challen (Addgene plasmid # 78254 ; http://n2t.net/addgene:78254 ; RRID:Addgene_78254)
For your References section:Reprogrammable CRISPR/Cas9-based system for inducing site-specific DNA methylation. McDonald JI, Celik H, Rois LE, Fishberger G, Fowler T, Rees R, Kramer A, Martens A, Edwards JR, Challen GA. Biol Open. 2016 May 11. pii: bio.019067. doi: 10.1242/bio.019067. 10.1242/bio.019067 PubMed 27170255