caa a22 4500 606175091 CHVBK 20210128100741.0 cr unu---uuuuu 210128e20151001xx s 000 0 eng 10.1007/s00018-015-1956-5 doi (NATIONALLICENCE)springer-10.1007/s00018-015-1956-5 Recent developments and clinical studies utilizing engineered zinc finger nuclease technology [Elektronische Daten] [Young-Il Jo, Hyongbum Kim, Suresh Ramakrishna] Efficient methods for creating targeted genetic modifications have long been sought for the investigation of gene function and the development of therapeutic modalities for various diseases, including genetic disorders. Although such modifications are possible using homologous recombination, the efficiency is extremely low. Zinc finger nucleases (ZFNs) are custom-designed artificial nucleases that make double-strand breaks at specific sequences, enabling efficient targeted genetic modifications such as corrections, additions, gene knockouts and structural variations. ZFNs are composed of two domains: (i) a DNA-binding domain comprised of zinc finger modules and (ii) the FokI nuclease domain that cleaves the DNA strand. Over 17years after ZFNs were initially developed, a number of improvements have been made. Here, we will review the developments and future perspectives of ZFN technology. For example, ZFN activity and specificity have been significantly enhanced by modifying the DNA-binding domain and FokI cleavage domain. Advances in culture methods, such as the application of a cold shock and the use of small molecules that affect ZFN stability, have also increased ZFN activity. Furthermore, ZFN-induced mutant cells can be enriched using episomal surrogate reporters. Additionally, we discuss several ongoing clinical studies that are based on ZFN-mediated genome editing in humans. These breakthroughs have substantially facilitated the use of ZFNs in research, medicine and biotechnology. Springer Basel, 2015 Farm animals nationallicence Pre-clinical trials nationallicence Programmable nucleases nationallicence Targeted genetic modifications nationallicence Therapeutic applications nationallicence ZFN architecture nationallicence ZFN delivery nationallicence ZFN modification nationallicence Jo Young-Il Brandeis University, Waltham, MA, USA aut Kim Hyongbum Department of Pharmacology and Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, Seoul, South Korea aut Ramakrishna Suresh Graduate School of Biomedical Science and Engineering, Hanyang University, Sungdong-gu, Seoul, South Korea aut Cellular and Molecular Life Sciences Springer Basel 72/20(2015-10-01), 3819-3830 1420-682X 72:20<3819 2015 72 18 https://doi.org/10.1007/s00018-015-1956-5 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 review-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00018-015-1956-5 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Jo Young-Il Brandeis University, Waltham, MA, USA aut NATIONALLICENCE 700 1- Kim Hyongbum Department of Pharmacology and Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, Seoul, South Korea aut NATIONALLICENCE 700 1- Ramakrishna Suresh Graduate School of Biomedical Science and Engineering, Hanyang University, Sungdong-gu, Seoul, South Korea aut NATIONALLICENCE 773 0- Cellular and Molecular Life Sciences Springer Basel 72/20(2015-10-01), 3819-3830 1420-682X 72:20<3819 2015 72 18