Home › Cell Biology › Genome Editing with CompoZr Custom Zinc Finger Nucleases (ZFNs)
Cell Biology JoVE (Open Access) Citable · DOI

Genome Editing with CompoZr Custom Zinc Finger Nucleases (ZFNs)

DOI: 10.3791/3304-v
What you'll learn
  • ✓ Design custom ZFNs targeting a user-defined genomic locus
  • ✓ Validate ZFN activity via Cel-1 nuclease assay
  • ✓ Deliver ZFNs into cells using nucleofection
  • ✓ Isolate and characterize clonal edited cell lines
Protocol

Biopharma Insights The CompoZr Custom Zinc-Finger Nuclease (ZFN) Service enables precise genome editing in any organism or cell line at any locus defined by the user. This article describes the process for the design, manufacture, validation and implementation of the CompoZr Custom ZFN Service.

Difficulty
advanced
Total time
~6-10 weeks from design to validated clones
Biosafety
BSL-2

Steps

1
Initiate the custom ZFN design process

Submit the target locus sequence and specifications to the CompoZr Custom ZFN Service to begin custom nuclease design.

▶ 01:36
2
Receive and validate custom ZFN production

Obtain the manufactured ZFN reagents and confirm their activity through the provider's validation pipeline.

▶ 03:23
3
Deliver validated ZFNs by nucleofection

Introduce the validated ZFN reagents into target cells using the nucleofection electroporation method.

▶ 04:16
4
Perform Cel-1 assay for editing detection

Use the Cel-1 mismatch cleavage assay to detect and confirm targeted genomic modifications in the cell population.

▶ 06:25
5
Isolate and characterize edited clones

Perform clonal isolation from the edited population and characterize individual clones for the desired modification.

▶ 07:09
6
Estimate ZFN activity from Cel-1 results

Quantify the Cel-1 assay band intensities to estimate the overall ZFN editing efficiency.

▶ 07:46
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