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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.

Add to Cart
$150 USD + shipping

Available to academics and nonprofits only.

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 PUF Assembly Scheme 1

Golden Gate assembly of PUF

Golden Gate PUF Assembly Scheme 2

Image from Abil et al. Journal of Biological Engineering 2014, 8:7 http://www.jbioleng.org/content/8/1/7

Kit Documentation

Icon PUF Assembly Kit.pdf (1.2 MB)

How 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 links to its kit plate map on the right. 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
Data cached @ 2017-11-24

Kit Plate Map - #1000000051

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