PUF Assembly Kit
(Kit #
1000000051
)
Depositing Lab:
Huimin Zhao
This 39 plasmid kit is used to assemble custom-made RNA-binding proteins for recognition of 8-base long targets within RNA. The kit is based on the Pumilio and FBF (PUF) homology domain of the full-length human Pumilio 1 protein.
The kit will be shipped to most countries as bacterial glycerol stocks, 96-well format, on dry ice. Samples should be frozen at -80°C immediately upon arrival. Scientists in countries that cannot receive dry ice shipments will receive a room temperature version of the kit that needs to be processed immediately upon arrival.
Original Publication
Modular assembly of designer PUF proteins for specific post-transcriptional regulation of endogenous RNA. Abil Z, Denard CA, Zhao H. J Biol Eng . 2014 Mar 1;8(1):7. doi: 10.1186/1754-1611-8-7. PubMed .
Description
This collection comprises 39 plasmids for assembly of a custom-made RNA-binding protein for recognition of an 8-base long target within an RNA of choice. The kit is based on the Pumilio and FBF (PUF) homology domain, which comprises amino acids 828-1176 of the full-length human Pumilio 1 protein. Eight modules corresponding to eight structural repeats of the PUF domain are assembled using the Golden Gate method. The collection comprises 4 variations of each module for the recognition of any of the 4 RNA bases. The kit also contains two receiving vectors, pET28-GG-PUF for Escherichia coli expression and pCMV-TTP (C147R)-GG-PUF for mammalian expression of PUF domain C-terminally fused to human tristetraprolin (TTP), a translational regulator.
Schematic of the main library components: 8 repeat modules, 4 variations of each module, and two receiving vectors
Golden Gate assembly of PUF
Image from Abil et al. Journal of Biological Engineering 2014, 8:7 http://www.jbioleng.org/content/8/1/7
Kit Documentation
PUF Assembly Kit.pdf 1.2Â MBHow to Cite this Kit
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which they were created, and include Addgene in the Materials and Methods of your future publications.
Modular assembly of designer PUF proteins for specific post-transcriptional regulation of endogenous RNA. Abil Z, Denard CA, Zhao H. J Biol Eng . 2014 Mar 1;8(1):7. doi: 10.1186/1754-1611-8-7. PubMed .
PUF Assembly Kit - #1000000051
- Resistance Color Key
Each circle corresponds to a specific antibiotic resistance in the kit plate map wells.
- Inventory
Searchable and sortable table of all plasmids in kit. The Well column lists the plasmid well location in its plate. The Plasmid column links to a plasmid's individual web page.
- Kit Plate Map
96-well plate map for plasmid layout. Hovering over a well reveals the plasmid name, while clicking on a well opens the plasmid page.
Resistance Color Key
| Ampicillin | |
| Chloramphenicol | |
| Kanamycin |
Inventory
| Well | Plasmid | Resistance |
|---|---|---|
| A / 1 | pCMV-TTP(C147R)-GG-PUF | Ampicillin |
| A / 2 | pChlr-pNEB193 | Chloramphenicol |
| A / 3 | pChlr-1SQ | Chloramphenicol |
| A / 4 | pChlr-1SE | Chloramphenicol |
| A / 5 | pChlr-1NQ | Chloramphenicol |
| A / 6 | pChlr-1SR | Chloramphenicol |
| A / 7 | pChlr-2CQ | Chloramphenicol |
| A / 8 | pChlr-2SE | Chloramphenicol |
| A / 9 | pChlr-2NQ | Chloramphenicol |
| A / 10 | pChlr-2SR | Chloramphenicol |
| A / 11 | pChlr-3CQ | Chloramphenicol |
| A / 12 | pChlr-3SE | Chloramphenicol |
| B / 1 | pChlr-3NQ | Chloramphenicol |
| B / 2 | pChlr-3SR | Chloramphenicol |
| B / 3 | pChlr-4CQ | Chloramphenicol |
| B / 4 | pChlr-4SE | Chloramphenicol |
| B / 5 | pChlr-4NQ | Chloramphenicol |
| B / 6 | pChlr-4SR | Chloramphenicol |
| B / 7 | pChlr-4CYQ | Chloramphenicol |
| B / 8 | pChlr-4SYE | Chloramphenicol |
| B / 9 | pChlr-4NYQ | Chloramphenicol |
| B / 10 | pChlr-4SYR | Chloramphenicol |
| B / 11 | pChlr-5CQ | Chloramphenicol |
| B / 12 | pChlr-5SE | Chloramphenicol |
| C / 1 | pChlr-5NQ | Chloramphenicol |
| C / 2 | pChlr-5SR | Chloramphenicol |
| C / 3 | pChlr-6CQ | Chloramphenicol |
| C / 4 | pChlr-6SE | Chloramphenicol |
| C / 5 | pChlr-6NQ | Chloramphenicol |
| C / 6 | pChlr-6SR | Chloramphenicol |
| C / 7 | pChlr-7CQ | Chloramphenicol |
| C / 8 | pChlr-7SE | Chloramphenicol |
| C / 9 | pChlr-7NQ | Chloramphenicol |
| C / 10 | pChlr-7SYR | Chloramphenicol |
| C / 11 | pChlr-8CQ | Chloramphenicol |
| C / 12 | pChlr-8SE | Chloramphenicol |
| D / 1 | pChlr-8NQ | Chloramphenicol |
| D / 2 | pChlr-8SR | Chloramphenicol |
| D / 3 | pET28-GG-PUF | Kanamycin |