Chapter title |
An Effective Method for Specific Gene Silencing in Escherichia coli Using Artificial Small RNA.
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Chapter number | 17 |
Book title |
RNA Scaffolds
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Published in |
Methods in molecular biology, January 2015
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DOI | 10.1007/978-1-4939-2730-2_17 |
Pubmed ID | |
Book ISBNs |
978-1-4939-2729-6, 978-1-4939-2730-2
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Authors |
Bak, Geunu, Choi, Jee Soo, Kim, Wonkyeong, Suk, Shinae, Lee, Younghoon, Geunu Bak, Jee Soo Choi, Wonkyeong Kim, Shinae Suk, Younghoon Lee |
Abstract |
Knockdown or silencing of a specific gene presents a powerful strategy for elucidating gene function in a variety of organisms. To date, efficient silencing methods have been established in eukaryotes, but not bacteria. In this chapter, an efficient and versatile gene silencing method using artificial small RNA (afsRNA) is described. For this purpose, target-recognizing sequences were introduced in specially designed RNA scaffolds to exist as single-stranded stretches in afsRNA. The translation initiation region of target genes was used as the sequence for afsRNA recognition, based on the theory that this site is usually highly accessible to ribosomes, and therefore, possibly, afsRNA.Two genes transcribed as monocistrons were tested with our protocol. Both genes were effectively silenced by their cognate afsRNAs. |
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