Helper-independent piggyBac plasmids for gene delivery approaches: Strategies for avoiding potential genotoxic effects
| العنوان: | Helper-independent piggyBac plasmids for gene delivery approaches: Strategies for avoiding potential genotoxic effects |
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| المؤلفون: | Ryuzo Yanagimachi, Kris Kawamoto, Ilko Stoytchev, Hideaki Yamashiro, James A. Dee, Stefan Moisyadi, Joseph M. Kaminski, Joel Marh, Kazuto Morozumi, Johann Urschitz, Craig J. Coates, Pawel Pelczar, Jesse B. Owens, Miyuri Kawasumi |
| المصدر: | Proceedings of the National Academy of Sciences. 107:8117-8122 |
| بيانات النشر: | Proceedings of the National Academy of Sciences, 2010. |
| سنة النشر: | 2010 |
| مصطلحات موضوعية: | Genetics, Transposable element, Multidisciplinary, Base Sequence, Genetic Vectors, Gene Transfer Techniques, Transposases, Genetic Therapy, Biological Sciences, Gene delivery, Biology, Cell Line, Viral vector, Insertional mutagenesis, Mice, Plasmid, Animals, Humans, Expression cassette, Gene, Transposase, DNA Damage, Plasmids |
| الوصف: | Efficient integration of functional genes is an essential prerequisite for successful gene delivery such as cell transfection, animal transgenesis, and gene therapy. Gene delivery strategies based on viral vectors are currently the most efficient. However, limited cargo capacity, host immune response, and the risk of insertional mutagenesis are limiting factors and of concern. Recently, several groups have used transposon-based approaches to deliver genes to a variety of cells. The piggyBac (pB) transposase in particular has been shown to be well suited for cell transfection and gene therapy approaches because of its flexibility for molecular modification, large cargo capacity, and high transposition activity. However, safety considerations regarding transposase gene insertions into host genomes have rarely been addressed. Here we report our results on engineering helper-independent pB plasmids. The single-plasmid gene delivery system carries both the piggyBac transposase (pBt) expression cassette as well as the transposon cargo flanked by terminal repeat element sequences. Improvements to the helper-independent structure were achieved by developing new plasmids in which the pBt gene is rendered inactive after excision of the transposon from the plasmid. As a consequence, potentially negative effects that may develop by the persistence of an active pBt gene posttransposition are eliminated. The results presented herein demonstrate that our helper-independent plasmids represent an important step in the development of safe and efficient gene delivery methods that should prove valuable in gene therapy and transgenic approaches. |
| تدمد: | 1091-6490 0027-8424 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::4d39430fd453b5a0c57fc8c148a0e325 https://doi.org/10.1073/pnas.1003674107 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....4d39430fd453b5a0c57fc8c148a0e325 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 10916490 00278424 |
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