PurposeTransient expression in mammalian cells of an "INT" construct_bearing P4_P6 (internally appended with three PP7 stem-loops), targeting the GLuc reporter. U6 promoter
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||68432||Standard format: Plasmid sent in bacteria as agar stab||1||$65|
This material is available to academics and nonprofits only.
Vector typeMammalian Expression, CRISPR, Synthetic Biology
Growth in Bacteria
Copy numberHigh Copy
Gene/Insert nameINT construct_bearing P4_P6 (internally appended with three PP7 stem-loops)
gRNA/shRNA sequencegRNA: GATCTAGATACGACTCACTAT
- Promoter human U6
- Cloning method Restriction Enzyme
- 5′ cloning site Unknown (unknown if destroyed)
- 3′ cloning site Unknown (unknown if destroyed)
- 5′ sequencing primer LKO.1 5
- 3′ sequencing primer M13R (Common Sequencing Primers)
sgRNA core targets the (ATCTAGATACGACTCACTAT) sequence in the Gluc reporter. Contains an internal "accessory domain," comprising the T. thermophilia P4-P6 domain appended with a casette of PP7 stem-loops.
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:pU6_(GLuc)_INT[P4-P6(3xPP7)] was a gift from John Rinn (Addgene plasmid # 68432 ; http://n2t.net/addgene:68432 ; RRID:Addgene_68432)
For your References section:Multiplexable, locus-specific targeting of long RNAs with CRISPR-Display. Shechner DM, Hacisuleyman E, Younger ST, Rinn JL. Nat Methods. 2015 Jul;12(7):664-70. doi: 10.1038/nmeth.3433. Epub 2015 Jun 1. 10.1038/nmeth.3433 PubMed 26030444