ArticlePloS one2013
Stability of zinc finger nuclease protein is enhanced by the proteasome inhibitor MG132.
Article in PloS one, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 18 papers.
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Who cites it
18 citing papers in PubMed, 22 citations in OpenAlex.
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- Diosgenin Promotes Bacterial Clearance in Bladder Epithelial Cells by Regulating Rab27b Expression: A Study Based on Virtual Screening and Experimental Validation.Infection and drug resistance · 2026Article
- ZNF821 as a potential biomarker associated with immune infiltration in pancreatic adenocarcinoma.Brazilian journal of medical and biological research = Revista brasileira de pesquisas medicas e biologicas · 2026Article
- Targeted DNA Demethylation: Vectors, Effectors and Perspectives.Biomedicines · 2023Review
- Article
- AnFrontiers in molecular neuroscience · 2021Article
- Genome editing in wheat microspores and haploid embryos mediated by delivery of ZFN proteins and cell-penetrating peptide complexes.Plant biotechnology journal · 2020Article
- In Vivo Delivery Systems for Therapeutic Genome Editing.International journal of molecular sciences · 2016Review
- Artificial zinc finger DNA binding domains: versatile tools for genome engineering and modulation of gene expression.Journal of cellular biochemistry · 2015Review
- Recent developments and clinical studies utilizing engineered zinc finger nuclease technology.Cellular and molecular life sciences : CMLS · 2015Review
- Covalent Incorporation of Trehalose within Hydrogels for Enhanced Long-Term Functional Stability and Controlled Release of Biomacromolecules.Advanced healthcare materials · 2015Article
- Rh D blood group conversion using transcription activator-like effector nucleases.Nature communications · 2015Article
- Integration of drug, protein, and gene delivery systems with regenerative medicine.Drug delivery and translational research · 2015Review
- Enhanced gene disruption by programmable nucleases delivered by a minicircle vector.Gene therapy · 2014Article
- Gene disruption by cell-penetrating peptide-mediated delivery of Cas9 protein and guide RNA.Genome research · 2014Article
- A guide to genome engineering with programmable nucleases.Nature reviews. Genetics · 2014Review
- Non-integrating gamma-retroviral vectors as a versatile tool for transient zinc-finger nuclease delivery.Scientific reports · 2014Article
- Functional analysis of a breast cancer-associated FGFR2 single nucleotide polymorphism using zinc finger mediated genome editing.PloS one · 2013Article
Corrections and comments
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Authors and funding
3 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundZinc finger nucleases (ZFNs) are powerful tools for gene therapy and genetic engineering. The characterization of ZFN protein stability and the development of simple methods to improve ZFN function would facilitate the application of this promising technology. However, the factors that affect ZFN protein stability and function are not yet clear. Here, we determined the stability and half-life of two ZFN proteins and examined the effect of MG132 (carbobenzoxyl-leucinyl-leucinyl-leucinal-Hl), a proteasome inhibitor, on ZFN-mediated gene modifications. METHODOLOGY/PRINCIPAL
findingsZFN proteins were expressed in 293T cells after transfection of ZFN-encoding plasmids. We studied two ZFN pairs: Z-224, which targets the CCR5 gene, and K-230, which targets a region 230 kbp upstream of CCR5. Western blotting after treatment with cycloheximide showed that the half-life of these ZFN proteins was around two hours. An immunoprecipitation assay revealed that the ZFN interacts with ubiquitin molecules and undergoes polyubiquitination in vivo. Western blotting showed that the addition of MG132, a proteasomal inhibitor, increased ZFN protein levels. Finally, a surrogate reporter assay and a T7E1 assay revealed that MG132 treatment enhanced ZFN-directed gene editing.
conclusionsTo our knowledge, this is the first study to investigate ZFN protein stability and to show that a small molecule can increase ZFN activity. Our protein stability study should lay the foundation for further improvement of ZFN technology; as a first step, the use of the small molecule MG132 can enhance the efficiency of ZFN-mediated gene editing.
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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.