ArticleBiochemistry2011
Adding fingers to an engineered zinc finger nuclease can reduce activity.
Article in Biochemistry, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 21 papers.
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Who cites it
21 citing papers in PubMed, 38 citations in OpenAlex.
- Xylt2 Knockout Eliminates O-Xylosylation and Enhances Bone Morphogenetic Protein 2 Production in Chinese Hamster Ovary Cells.Biotechnology journal · 2026Article
- Current trends in application of CRISPR/Cas9 in gene editing and diagnostics in Neglected tropical diseases (NTDs).Molecular biology reports · 2025Review
- Prokaryotic Argonaute Proteins as a Tool for Biotechnology.Molecular biology · 2022Article
- Comparison of the Feasibility, Efficiency, and Safety of Genome Editing Technologies.International journal of molecular sciences · 2021Review
- Protein Delivery of Cell-Penetrating Zinc-Finger Activators Stimulates Latent HIV-1-Infected Cells.Molecular therapy. Methods & clinical development · 2020Article
- Balance between asymmetry and abundance in multi-domain DNA-binding proteins may regulate the kinetics of their binding to DNA.PLoS computational biology · 2020Article
- Gene Editing and Alzheimer's Disease: Is There Light at the End of the Tunnel?Frontiers in genome editing · 2020Review
- Genome editing of oncogenes with ZFNs and TALENs: caveats in nuclease design.Cancer cell international · 2018Article
- Balancing between affinity and speed in target DNA search by zinc-finger proteins via modulation of dynamic conformational ensemble.Proceedings of the National Academy of Sciences of the United States of America · 2015Article
- The I-TevI nuclease and linker domains contribute to the specificity of monomeric TALENs.G3 (Bethesda, Md.) · 2014Article
- Highly active zinc-finger nucleases by extended modular assembly.Genome research · 2013Article
- Using defined finger-finger interfaces as units of assembly for constructing zinc-finger nucleases.Nucleic acids research · 2013Article
- Repeatable construction method for engineered zinc finger nuclease based on overlap extension PCR and TA-cloning.PloS one · 2013Article
- Monomeric site-specific nucleases for genome editing.Proceedings of the National Academy of Sciences of the United States of America · 2012Article
- Gene targeting to the ROSA26 locus directed by engineered zinc finger nucleases.Nucleic acids research · 2012Article
- Zinc Finger Nucleases: Tailor-made for Gene Therapy.Drugs of the future · 2012Article
- Versatile and efficient genome editing in human cells by combining zinc-finger nucleases with adeno-associated viral vectors.Human gene therapy · 2012Article
- Zinc-finger nucleases: how to play two good hands.Nature methods · 2011Article
- Revealing off-target cleavage specificities of zinc-finger nucleases by in vitro selection.Nature methods · 2011Article
- Selection-independent generation of gene knockout mouse embryonic stem cells using zinc-finger nucleases.PloS one · 2011Article
Corrections and comments
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Authors and funding
7 authors at 2 institutions in 2 countries.
Funding
Abstract
Zinc finger nucleases (ZFNs) have been used to direct precise modifications of the genetic information in living cells at high efficiency. An important consideration in the design of ZFNs is the number of zinc fingers that are required for efficient and specific cleavage. We examined dimeric ZFNs composed of [1]+[1], [2]+[2], [3]+[3], [4]+[4], [5]+[5], and [6]+[6] zinc fingers, targeting 6, 12, 18, 24, 30, and 36 bp, respectively. We found that [1]+[1] and [2]+[2] fingers supported neither in vitro cleavage nor single-strand annealing in a cell-based recombination assay. An optimal ZFN activity was observed for [3]+[3] and [4]+[4] fingers. Surprisingly, [5]+[5] and [6]+[6] fingers exhibited significantly reduced activity. While the extra fingers were not found to dramatically increase toxicity, directly inhibit recombination, or perturb the ZFN target site, we demonstrate the ability of subsets of three fingers in six-finger arrays to bind independently to regions of the target site, possibly explaining the decrease in activity. These results have important implications for the design of new ZFNs, as they show that in some cases an excess of fingers may actually negatively affect the performance of engineered multifinger proteins. Maximal ZFN activity will require an optimization of both DNA binding affinity and specificity.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.