Cited by 129CRISPR and Genetic EngineeringRNA regulation and diseaseRNA Interference and Gene Delivery

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Design, engineering, and characterization of zinc finger nucleases

Mani, Mala, Kandavelou, Karthikeyan, Dy, Fei Jamie, Durai, Sundar, Chandrasegaran, Srinivasan·2005·Biochemical and Biophysical Research Communications

DOI 10.1016/j.bbrc.2005.07.089

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Abstract

Zinc finger nuclease (ZFN)-mediated gene targeting is rapidly becoming a powerful tool for "gene editing" and "directed mutagenesis" of plant and mammalian genomes including the human genome. ZFN-mediated gene targeting provides molecular biologists with the ability to site-specifically manipulate and permanently modify plant and mammalian genomes. Facile production of ZFNs and rapid characterization of their in vitro sequence-specific cleavage properties are a pre-requisite before ZFN-mediated gene targeting can become an efficient and effective practical tool for widespread use in biotechnology. Here, we report the design, engineering, and rapid in vitro characterization of ZFNs that target specific endogenous sequences within two mouse genes (mTYR and mCFTR), and two human genes (hCCR5 and hDMPK), respectively. These engineered ZFNs recognize their respective cognate DNA sites encoded in a plasmid substrate in a sequence-specific manner and, as expected, they induce a double-strand break at the chosen target site.

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Authors: Mani, M., Kandavelou, K., Dy, F. J., Durai, S., & Chandrasegaran, S.
Affiliations: Johns Hopkins University (US)
Corresponding author: Chandrasegaran, Srinivasan
ORCID: Dy, Fei Jamie
Venue: Biochemical and Biophysical Research Communications
Publisher: Elsevier BV
Published: 2005-09
Volume: 335
Issue: 2
Pages: 447-457
ISSN: 0006-291X (ISSN-L)
DOI: 10.1016/j.bbrc.2005.07.089
OpenAlex: W1972389078
Cited by: 129
References: 39
Citations by year: 2026: 1 · 2025: 5 · 2024: 6 · 2023: 4 · 2022: 4 · 2021: 5 · 2020: 3 · 2019: 4 · 2018: 4 · 2017: 2 · 2016: 7 · 2015: 7 · 2014: 6 · 2013: 7 · 2012: 8
FWCI: 3.36
Type: journal article
Language: en
Open access: closed
Licence: www.elsevier.com/tdm/userlicense/1.0/ (tdm)
Locations: Biochemical and Biophysical Research Communications (publishedVersion)
PubMed (publishedVersion)
Funders: National Institutes of Health
National Institute of General Medical Sciences (R01GM053923, GM 53923)
National Institute of Allergy and Infectious Diseases (P30AI042855)
Topics: Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA regulation and disease
Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
Keywords: zinc finger nucleases, gene targeting, genome editing, directed mutagenesis, sequence-specific cleavage, DNA double-strand breaks, tyrosinase, CFTR, CCR5, DMPK
MeSH terms: Amino Acid Sequence, Animals, Base Sequence, DNA, DNA Damage, Endoribonucleases, Genetic Techniques, Humans, Molecular Sequence Data, Plasmids, Rabbits, Recombination, Genetic, Transcription, Genetic, Protein Biosynthesis, Blotting, Western, Protein Engineering, Genome, Human, Polymerase Chain Reaction, Mutagenesis, Site-Directed, Zinc Fingers, Genome, Protein Serine-Threonine Kinases, Protein Structure, Tertiary, Genome, Plant, Receptors, CCR5, Amino Acid Motifs, Mice, Myotonin-Protein Kinase

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Design, engineering, and characterization of zinc finger nucleases

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