Using defined finger–finger interfaces as units of assembly for constructing zinc-finger nucleases
| العنوان: | Using defined finger–finger interfaces as units of assembly for constructing zinc-finger nucleases |
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| المؤلفون: | Gary D. Stormo, Cong Zhu, Abirami Lakshmanan, Amy L. Rayla, Ankit Gupta, Benjamin Dake, Victoria L. Hall, Scot A. Wolfe, Charlotte Kuperwasser, Ryan G. Christensen |
| المصدر: | Nucleic Acids Research |
| بيانات النشر: | Oxford University Press, 2013. |
| سنة النشر: | 2013 |
| مصطلحات موضوعية: | Recombinant Fusion Proteins, Computational biology, Biology, Protein Engineering, Genome, Domain (software engineering), Genome engineering, Image stitching, 03 medical and health sciences, 0302 clinical medicine, Genetics, Animals, Humans, Deoxyribonucleases, Type II Site-Specific, Zebrafish, 030304 developmental biology, Zinc finger, 0303 health sciences, Binding Sites, Deoxyribonucleases, Nucleic Acid Enzymes, Nucleotides, fungi, Zinc Fingers, DNA, Zinc finger nuclease, DNA metabolism, body regions, HEK293 Cells, 030217 neurology & neurosurgery |
| الوصف: | Zinc-finger nucleases (ZFNs) have been used for genome engineering in a wide variety of organisms; however, it remains challenging to design effective ZFNs for many genomic sequences using publicly available zinc-finger modules. This limitation is in part because of potential finger-finger incompatibility generated on assembly of modules into zinc-finger arrays (ZFAs). Herein, we describe the validation of a new set of two-finger modules that can be used for building ZFAs via conventional assembly methods or a new strategy-finger stitching-that increases the diversity of genomic sequences targetable by ZFNs. Instead of assembling ZFAs based on units of the zinc-finger structural domain, our finger stitching method uses units that span the finger-finger interface to ensure compatibility of neighbouring recognition helices. We tested this approach by generating and characterizing eight ZFAs, and we found their DNA-binding specificities reflected the specificities of the component modules used in their construction. Four pairs of ZFNs incorporating these ZFAs generated targeted lesions in vivo, demonstrating that stitching yields ZFAs with robust recognition properties. |
| اللغة: | English |
| تدمد: | 1362-4962 0305-1048 |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::0733d7171f07a4dd64b3a9742f042fd3Test http://europepmc.org/articles/PMC3575815Test |
| حقوق: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....0733d7171f07a4dd64b3a9742f042fd3 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 13624962 03051048 |
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