Construction of a guide-RNA for site-directed RNA mutagenesis utilising intracellular A-to-I RNA editing
| Autor: | Azusa Nishitarumizu, Masatora Fukuda, Kanako Nose, Hiroyuki Nakagawa, Ryoma Noguchi, Hiromitsu Umeno |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Adenosine Adenosine Deaminase Protein domain Intracellular Space Mutagenesis (molecular biology technique) Computational biology Biology Article 03 medical and health sciences chemistry.chemical_compound Protein Domains Humans Guide RNA Codon Genetics Multidisciplinary Base Sequence HEK 293 cells RNA RNA-Binding Proteins Inosine 030104 developmental biology HEK293 Cells chemistry RNA editing ADAR Mutagenesis Site-Directed RNA Editing DNA RNA Guide Kinetoplastida |
| Zdroj: | Scientific Reports |
| ISSN: | 2045-2322 |
| Popis: | As an alternative to DNA mutagenesis, RNA mutagenesis can potentially become a powerful gene-regulation method for fundamental research and applied life sciences. Adenosine-to-inosine (A-to-I) RNA editing alters genetic information at the transcript level and is an important biological process that is commonly conserved in metazoans. Therefore, a versatile RNA-mutagenesis method can be achieved by utilising the intracellular RNA-editing mechanism. Here, we report novel guide RNAs capable of inducing A-to-I mutations by guiding the editing enzyme, human adenosine deaminase acting on RNA (ADAR). These guide RNAs successfully introduced A-to-I mutations into the target-site, which was determined by the reprogrammable antisense region. In ADAR2-over expressing cells, site-directed RNA editing could also be performed by simply introducing the guide RNA. Our guide RNA framework provides basic insights into establishing a generally applicable RNA-mutagenesis method. |
| Databáze: | OpenAIRE |
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