A simple method for construction of artificial microRNA vector in plant
2014 (English)In: Biotechnology letters, ISSN 0141-5492, E-ISSN 1573-6776, Vol. 36, no 10, 2117-2123 p.Article in journal (Refereed) Published
Artificial microRNA (amiRNA) is a powerful tool for silencing genes in many plant species. Here we provide an easy method to construct amiRNA vectors that reinvents the Golden Gate cloning approach and features a novel system called top speed amiRNA construction (TAC). This speedy approach accomplishes one restriction-ligation step in only 5 min, allowing easy and high-throughput vector construction. Three primers were annealed to be a specific adaptor, then digested and ligated on our novel vector pTAC. Importantly, this method allows the recombined amiRNA constructs to maintain the precursor of osa-miR528 with exception of the desired amiRNA/amiRNA* sequences. Using this method, our results showed the expected decrease of targeted genes in Nicotiana benthamiana and Oryza sativa.
Place, publisher, year, edition, pages
2014. Vol. 36, no 10, 2117-2123 p.
Artificial microRNA, Gene silencing, Golden gate cloning, High-throughput construction, Nicotiana benthamiana, Osa-miR528
Other Biological Topics
IdentifiersURN: urn:nbn:se:kth:diva-153841DOI: 10.1007/s10529-014-1570-xISI: 000341486600025ScopusID: 2-s2.0-84906948235OAI: oai:DiVA.org:kth-153841DiVA: diva2:755058
QC 201410132014-10-132014-10-092014-10-13Bibliographically approved