Ochiai Hiroshi, Yamamoto Takashi
PRESTO, JST, Higashi-Hiroshima, 739-8530, Japan.
Department of Mathematical and Life Sciences, Graduate School of Science, Hiroshima University, Higashi-Hiroshima, 739-8530, Japan.
Methods Mol Biol. 2017;1630:1-24. doi: 10.1007/978-1-4939-7128-2_1.
Zinc-finger nucleases (ZFNs) are programmable nucleases that have opened the door to the genome editing era. The construction of ZFNs recognizing a target sequence of interest is laborious, and has not been widely used recently. However, key ZFN patents are expiring over the next 2-4 years, enabling a wide range of deployments for clinical and industrial applications. This article introduces a ZFN construction protocol that uses bacterial one-hybrid (B1H) selection and single-stranded annealing (SSA) assay.
锌指核酸酶（ZFNs）是可编程核酸酶，开启了基因组编辑时代的大门。构建识别目标序列的锌指核酸酶过程繁琐，近年来未得到广泛应用。然而，关键的锌指核酸酶专利将在未来2至4年内到期，这将使得其在临床和工业应用中有广泛的部署。本文介绍了一种利用细菌单杂交（B1H）筛选和单链退火（SSA）检测的锌指核酸酶构建方案。
