PurposeExpression of increased fidelity SpCas9-HF1-plus in bacterial cells. Cleaves both 20nt and 5'-extended 21nt spacer containing sgRNAs with close to the same fidelity as SpCas9-HF1.
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||126775||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
- Total vector size (bp) 10734
Vector typeBacterial Expression, CRISPR
Growth in Bacteria
Copy numberHigh Copy
Insert Size (bp)5436
MutationN497A; Q695A; Q926A; amino acids 1005-1013 replaced with two glycine
- Promoter T7
/ Fusion Proteins
- 3xFLAG (N terminal on insert)
- NLS (N terminal on insert)
- NLS (C terminal on insert)
- MBP (N terminal on backbone)
- 6xHis (N terminal on backbone)
- Cloning method Restriction Enzyme
- 5′ cloning site BcuI (not destroyed)
- 3′ cloning site NotI (not destroyed)
- 5′ sequencing primer CGGCGTACTGGTCGCCGATC
- 3′ sequencing primer TGCTAGTTATTGCTCAGCGG (Common Sequencing Primers)
Terms and Licenses
- Not Available to Industry
SpCas9-HF1-plus cleaves both 20nt and 5'-extended 21nt spacer containing sgRNAs. Shows almost the same fidelity and target selectivity as its parental increased fidelity variant, SpCas9-HF1.
For detailed information and plasmid usage, please see the associated publication.
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:pET-FLAG-SpCas9-HF1-plus was a gift from Ervin Welker (Addgene plasmid # 126775 ; http://n2t.net/addgene:126775 ; RRID:Addgene_126775)
For your References section:Blackjack mutations improve the on-target activities of increased fidelity variants of SpCas9 with 5'G-extended sgRNAs. Kulcsar PI, Talas A, Toth E, Nyeste A, Ligeti Z, Welker Z, Welker E. Nat Commun. 2020 Mar 6;11(1):1223. doi: 10.1038/s41467-020-15021-5. 10.1038/s41467-020-15021-5 PubMed 32144253