Construction and Evaluation of Zinc Finger Nucleases.

Hiroshi Ochiai, Takashi Yamamoto

Methods in molecular biology (Clifton, N.J.) 2023

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Zinc finger nucleases (ZFNs) are programmable nucleases that have contributed significantly to past genome-editing research. They are now utilized much less owing to the advent of transcription activator-like effector nucleases (TALENs) and the clustered regularly interspaced short palindromic repeats and CRISPR-associated protein system (CRISPR-Cas). These new methods allow for easier generation of reagents that target genomic sequences of interest and are less labor-intensive than ZFNs at targeting desired sequences. However, fundamental ZFN patents have expired, enabling a wide range of their distribution for clinical and industrial applications. This article introduces a ZFN construction protocol that uses bacterial one-hybrid (B1H) selection and single-strand annealing (SSA) assay. © 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.

Citation

Hiroshi Ochiai, Takashi Yamamoto. Construction and Evaluation of Zinc Finger Nucleases. Methods in molecular biology (Clifton, N.J.). 2023;2637:1-25