CompoZr™ Custom ZFN Service

Benefits

• Rapid design, assembly, and validation of a ZFN pair targeting your gene of interest
• Rapid and permanent disruption of, or integration into, any genomic loci

Cell Line Engineering Made Easy
CompoZr Zinc finger nucleases (ZFNs) are a class of engineered DNA-binding proteins that facilitate targeted editing of the genome by creating double-strand breaks in DNA at user specified locations. Researchers use ZFNs to create modified cell lines with gene deletions, gene integrations, and gene corrections. The CompoZr Custom ZFN Service provides researchers with a single pair of ZFNs that have been specifically designed, assembled, and validated to edit their target gene of interest.

Initiating a CompoZr Custom ZFN Project
The following information is required to initiate a CompoZr Custom ZFN Project:

• Organism of Interest
  • Human
  • Rat
  • Mouse
  • Hamster

• Goal of Your Gene Editing Project
  • Gene Deletion
  • Gene Integration
  • Gene Correction

• Target Gene Information
  • Gene Name
  • Gene ID, Accession #
  • Known Gene Homologs, Pseudogenes, Isoforms

• Intended Cell Line Usage
  • Basic Research
  • Target Validation
  • Cell-Based Screening
  • Cell Line Optimization

ZFN Candidate Design
Once an order is placed, Sigma's Bio-Informatics team will use the Sangamo SuperFinder algorithm to design ZFN candidates targeting the gene region of interest. The SuperFinder algorithm, continuously updated by Sangamo Biosciences and trained on feedback from wet lab results, has been designed to eliminate ZFN candidates of low specificity and binding efficacy.

ZFN Candidate Assembly
ZFN candidates designated by the SuperFinder algorithm will be assembled in the lab. First, identified Zinc Finger Protein (ZFP) modules from the proprietary Sangamo ZFN archive will be assembled to have a target-binding specificity of at least 24 bp. Next, a DNA-cleaving Nuclease domain will be added to each target specific ZFP to create assembled ZFN candidates.

ZFN Candidate Validation
The final step of the CompoZr ZFN-by-Design service involves validation of ZFN candidates in a proxy cell line specific to the organism of interest: Human, Mouse, Hamster, or Rat. To validate each ZFN, Sigma transfects the lead ZFNs into a proxy cell line and uses a mismatch sensitive nuclease assay to identify ZFNs that cleave the target site as expected. The ZFN pair with the greatest DNA-cleavage efficiency, as indicated by the mismatch assay, will be delivered to the customer.

CompoZr Custom ZFN Deliverables
Each Custom ZFN Project is managed by a dedicated Project Manager as your project moves through the three elements of the Custom ZFN service; ZFN Design, Assembly, and Validation. Deliverables include the following:

• Best Performing ZFN Pair
  • 10 Aliquots of Ready-to-Deliver ZFN Pair in mRNA form
  • ZFN Pair in Plasmid Form

• Forward and Reverse Primers that allow for determination of rate of mutation and for screening of individual clones
• Positive Control DNA
  • Used to determine a baseline cutting efficiency

Ordering Information
Please fill out the CompoZr ZFN Order Form and a Sigma® representative will contact you in regards to your project.

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