Efficient disruption of endogenous Bombyx gene by TAL effector nucleases |
| |
| Authors: | Suresh Sajwan Yoko Takasu Toshiki Tamura Keiro Uchino Hideki Sezutsu Michal Zurovec |
| |
| Affiliation: | 1. Advanced Clinical Research Center, Institute for Neurological Disorders, Fukushima and Kanagawa, Japan;2. Department of Pediatrics, Osaka University Graduate School of Medicine, Osaka, Japan;3. Department of Pediatrics, Akita University Graduate School of Medicine, Akita, Japan;4. Ohta General Hospital, Ohta Nishinouchi Hospital, Fukushima, Japan;5. Department of Neurology, Dokkyo University School of Medicine, Tochigi, Japan;6. Division of Child Neurology, Institute Neurological Sciences, Tottori University Faculty of Medicine, Tottori, Japan;7. Department of Gene Therapy, Institute of DNA Medicine, Jikei University School of Medicine, Tokyo, Japan;8. Department of Pediatrics, Jikei University School of Medicine, Tokyo, Japan;9. Jikei University School of Medicine, Tokyo, Japan;1. Department of Urology, Faculty of Medical Sciences, University of Fukui, Fukui, Japan;2. Molecular Imaging Center, National Institute of Radiological Sciences, 4-9-1, Anagawa, Inage, Chiba 263–8555, Japan;3. Graduate School of Pharmaceutical Sciences, The University of Tokyo, Tokyo, Japan;4. Sugiyama Laboratory, RIKEN Innovation Center, Research Cluster for Innovation, RIKEN, Kanagawa, Japan |
| |
| Abstract: | Engineered nucleases are proteins that are able to cleave DNA at specified sites in the genome. These proteins have recently been used for gene targeting in a number of organisms. We showed earlier that zinc finger nucleases (ZFNs) can be used for generating gene-specific mutations in Bombyx mori by an error-prone DNA repair process of non-homologous end joining (NHEJ). Here we test the utility of another type of chimeric nuclease based on bacterial TAL effector proteins in order to induce targeted mutations in silkworm DNA. We designed three TAL effector nucleases (TALENs) against the genomic locus BmBLOS2, previously targeted by ZFNs. All three TALENs were able to induce mutations in silkworm germline cells suggesting a higher success rate of this type of chimeric enzyme. The efficiency of two of the tested TALENs was slightly higher than of the successful ZFN used previously. Simple design, high frequency of candidate targeting sites and comparable efficiency of induction of NHEJ mutations make TALENs an important alternative to ZFNs. |
| |
| Keywords: | |
| 本文献已被 ScienceDirect 等数据库收录！ |
|
