PurposeTransient expression of the "TOP1" construct, targeting the GLuc reporter, in mammalian cells. U1Pro/SmBox/3' Box expression backbone.
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||68437||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 nameTOP1 construct
gRNA/shRNA sequencegRNA: GATCTAGATACGACTCACTAT
- Promoter human U1
- Cloning method Ligation Independent Cloning
- 5′ sequencing primer M13F
- 3′ sequencing primer M13R (Common Sequencing Primers)
sgRNA core targets the (ATCTAGATACGACTCACTAT) sequence in the Gluc reporter. Contains a 3' -"accessory domain," comprising the T. thermophilia P4-P6 domain appended with a casette of PP7 stem-loops. Trascription termination is by the U1 snRNA 3' Box system, preceeded by the U2 snRNA sm Box.
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:pU1/Sm/3' Box_(GLuc)_TOP1 was a gift from John Rinn (Addgene plasmid # 68437 ; http://n2t.net/addgene:68437 ; RRID:Addgene_68437)
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