CompoZr® Custom ZFN Service
Unlimited Genomic Engineering Possibilities in any Species or Cell Line
CompoZr Zinc Finger Nucleases (ZFNs) are a class of engineered DNA-binding proteins that facilitate highly specific targeted editing of any genome by creating double-stranded breaks in DNA at user specified locations. You can use ZFNs to create your choice of stable genetically engineered cell lines or organisms with gene deletions, gene integrations and gene modifications. The CompoZr Custom ZFN Service provides researchers with a functionally validated pair of ZFNs that have been custom designed and assembled to edit a target gene in a highly specific manner. Modify any gene in any biological system!
The following graphic highlights the abundance of genetic modifications that may be conducted using the Custom ZFN Service in any species or cell type.
||CompoZr Custom Zinc Finger Nuclease - with ZFN Plasmid only
|CompoZr Custom Zinc Finger Nuclease - with ZFN Plasmid and mRNA
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Watch the JOVE Video on Genome Editing with ZFNs
Now Available – Donor Design Service
The Donor Design Service can be used in conjunction with functionally validated CompoZr ZFNs to perform precise genome modifications in a variety of organisms or cell lines. Researcher-defined specifications are used to design in-silico donor sequences in plasmid DNA or single-stranded DNA (oligo) format. Once the donor sequence is received researchers should have the donor synthesized in preparation for targeted integration with the appropriate CompoZr ZFNs.
- Ability to conduct genetic modifications in any cell line or organism of your choice
- Quick, permanent and highly specific modification of any endogenous genome
- Edits induced through a single transfection
- Screen for gene disruption in as little as 48 hours
- Single or biallelic edits occur in 1-20% of clone population
- No antibiotic selection required for screening
- Exceptional mutant rate in founder animals with high percentage of germline transmission
- Rapid design, assembly and validation by Sigma of a ZFN pair targeting your gene of interest
- Immediately transfect your cell line once Custom ZFNs are received
- Receive ZFNs as plasmid or as plasmid plus ZFN mRNA format
- All Custom ZFN projects receive technical support from Sigma ZFN Technical Liaison
- CompoZr ZFN technology provides for targeted single locus gene modification
- ZFN activity an non-specific sites are negligible, the following publications demonstrate ZFN specificity
- Click here to view more zinc finger nuclease publications
CompoZr Custom ZFN Components
For increased access to ZFN technology Sigma Life Science provides Custom ZFNs in two formats. The first format includes ZFNs as plasmids only, for an additional price ZFNs are provided as plasmids plus 10 aliquots of ready-to-use ZFN mRNA.
- Plasmid DNA for each ZFN in the pair
- Forward and reverse primers for screening the DNA locus for mutation
- Genomic DNA as a positive control for PCR (from validated samples where the target gene has been mutated using the supplied CompoZr Custom ZFN)
- 10 aliquots of ready-to-deliver ZFN mRNA (optional component)
Researchers with human, mouse, rat and CHO ZFN projects receive the best performing pair of ZFNs after functional validation. Projects that are conducted in any other species are functionally validated in a yeast based assay and researchers will receive between 1-3 of the best performing ZFN pairs.
ZFN Custom Design Process
ZFN Candidate Design
Once an order is placed, Sigma's bioinformatics team will use the Sangamo proprietary algorithm to design ZFN candidates targeting the gene region of interest. This proprietary algorithm, continuously updated by Sangamo Biosciences and trained on feedback from validation testing, has been designed to yield ZFNs with high binding affinity to their specific target site.
ZFN Candidate Design Approval
Once the ZFN candidates have been designed, the selected candidates are sent to you, the customer, for verification. Only after the ZFN designs have been approved by you, do we start production.
ZFN Candidate Assembly
ZFN candidates approved by you will be assembled by the CompoZr ZFN Production Team. Using the Zinc Finger Protein (ZFP) modules from the proprietary Sangamo ZFN archive the production team will assemble sequence-specific ZFPs and attach the FokI nuclease domain creating a ZFN. Sigma-Aldrich produced ZFNs will have a 24-36 base pair specific target site.
ZFN Candidate Validation
The final step of the Custom CompoZr ZFN service involves validation of ZFN candidates.
- If the desired modification is in the human, mouse, rat or hamster genome, cell lines from the specific organism will be used for validation. Each ZFN pair constructed is transfected into the cells, and a mismatch sensitive assay is used to identify the functional ZFN pair. Cell line validated ZFNs require 10 weeks for production and validation, starting at your design approval.*
- If the desired modification is in the genome of any other organism, a yeast-based proxy system will be used, that accurately reflects ZFN activity in several cell types. Each ZFN pair constructed is transfected into the yeast-based proxy cells and a mismatch sensitive assay is used to identify the functional ZFN pair. Yeast-based proxy validated ZFNs require 12 weeks for production and validation, starting at your design approval.*
*Assumes successful validation of at least one pair of ZFNs from the initial design. Our on time delivery rate is > 80%.
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