PurposeMiddle Entry clone for Gateway containing a zebrafish codon-optimized Cas9 flanked by 2 NLS and followed by inframe GFP. Cas9 and GFP are separated by the sequence of a T2A self-cleaving peptide.
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||63155||Standard format: Plasmid sent in bacteria as agar stab||1||$65|
This material is available to academics and nonprofits only.
Vector backbonepME-MCS (Tol2Kit #237)
- Backbone size w/o insert (bp) 2765
- Total vector size (bp) 7685
Vector typeCRISPR ; Tol2 Middle Entry vector for Gateway
Growth in Bacteria
SpeciesD. rerio (zebrafish)
Insert Size (bp)4984
- Cloning method Restriction Enzyme
- 5′ cloning site SalI (not destroyed)
- 3′ cloning site NotI (not destroyed)
- 5′ sequencing primer CCTACTCAGGAGAGCGTTCA
- 3′ sequencing primer CAGGAAACAGCTATGACCAT (Common Sequencing Primers)
A portion of this plasmid was derived from a plasmid made bythe Chen laboratory (Jao et al., PNAS 2013).
Terms and Licenses
Articles Citing this Plasmid
Reference for zebrafish codon-optimized Cas9:
Jao, L.E., Wente, S.R., and Chen, W. (2013). Efficient multiplex biallelic zebrafish genome editing using a CRISPR nuclease system. Proc Natl Acad Sci U S A 110, 13904-13909.
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:pME-Cas9-T2A-GFP was a gift from Leonard Zon (Addgene plasmid # 63155 ; http://n2t.net/addgene:63155 ; RRID:Addgene_63155)
For your References section:A CRISPR/Cas9 Vector System for Tissue-Specific Gene Disruption in Zebrafish. Ablain J, Durand EM, Yang S, Zhou Y, Zon LI. Dev Cell. 2015 Mar 4. pii: S1534-5807(15)00075-1. doi: 10.1016/j.devcel.2015.01.032. 10.1016/j.devcel.2015.01.032 PubMed 25752963