RNA Interference by Production of Short Hairpin dsRNA in ES Cells, Their Differentiated Derivatives, and in Somatic Cell Lines
Abstract
dsRNA of several hundred nucleotides in length is effective at interfering with gene expression in mouse oocytes, pre-implantation embryos, and embryonic stem (ES) cells but is not as efficient in differentiated cell lines. Here we describe a method to achieve RNA interference in totipotent and differentiated ES cells together with a wide range of other mammalian cell types that is both simple and efficient. It utilizes a linearized plasmid that directs the expression of a hairpin RNA with a 22-nucleotide-paired region. This molecule has a 13-nucleotide 5′ overhang that would be subject to capping on its 5′ phosphoryl group and thus differs from the ideal structure suggested for effective small interfering RNAs. Thus, it appears either that the structure of small inhibitory RNA molecules may not need to be as precise as previously thought or that such a transcript is efficiently processed to a form that is effective in interfering with gene expression.
Additional Information
CC BY-NC-ND license. Received 23 October 2002; accepted 17 January 2003. We would like to thank Dr Simon Green from Cyclacel, Ltd. for inspiring discussion that led us to perform this work. M.Z.-G. and D.M.G. are grateful to Cancer Research UK (London, UK) for its contribution towards the support of this work through a grant awarded through Cancer Research Ventures (London, UK). J.B.G. and F.W. contributed equally to this work.Attached Files
Published - 03344st02.pdf
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Additional details
- Eprint ID
- 94826
- Resolver ID
- CaltechAUTHORS:20190419-105738699
- Cancer Research UK
- Created
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2019-04-23Created from EPrint's datestamp field
- Updated
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2021-11-16Created from EPrint's last_modified field