PurposegRNA expression vector (with mKate2)
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||73501||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
- Total vector size (bp) 9596
Vector typeMammalian Expression, Lentiviral, CRISPR
Growth in Bacteria
Growth Strain(s)NEB Stable
Copy numberHigh Copy
gRNA/shRNA sequenceScramble gRNA sequence with BsmBI cloning sites
SpeciesH. sapiens (human), Synthetic
- Promoter CAG
/ Fusion Protein
- NLS (N terminal on insert)
- Cloning method Restriction Enzyme
- 5′ cloning site EcoRI (not destroyed)
- 3′ cloning site EcoRI (not destroyed)
- 5′ sequencing primer CCTCTGCTAACCATGTTCATGC
- 3′ sequencing primer GATCTACCACATTTGTAGAG (Common Sequencing Primers)
Due to the large size of this gRNA expression vector it does not produce high-tire lentivirus. Hence we do not recommend using this plasmid backbone for lentivral preparation. This gRNA-expression plasmid is best suited for plasmid transfections and generation of stable clones.
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:pgRNA-CKB was a gift from Bruce Conklin (Addgene plasmid # 73501 ; http://n2t.net/addgene:73501 ; RRID:Addgene_73501)
For your References section:CRISPR Interference Efficiently Induces Specific and Reversible Gene Silencing in Human iPSCs. Mandegar MA, Huebsch N, Frolov EB, Shin E, Truong A, Olvera MP, Chan AH, Miyaoka Y, Holmes K, Spencer CI, Judge LM, Gordon DE, Eskildsen TV, Villalta JE, Horlbeck MA, Gilbert LA, Krogan NJ, Sheikh SP, Weissman JS, Qi LS, So PL, Conklin BR. Cell Stem Cell. 2016 Apr 7;18(4):541-53. doi: 10.1016/j.stem.2016.01.022. Epub 2016 Mar 10. 10.1016/j.stem.2016.01.022 PubMed 26971820