Linked Life Data

20569207

Source:http://linkedlifedata.com/resource/pubmed/id/20569207

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
4
pubmed:dateCreated
2010-6-23
pubmed:abstractText
The cleavage of double-stranded RNA (dsRNA) molecules by Dicer is a critical step in silencing genes by the RNA induced silencing complex (RISC). The development of Dicer substrates as nucleic acid-based therapeutics brings about a need to rapidly evaluate chemically modified RNA molecules for their ability to be processed by Dicer. This study outlines a quantitative fluorescence quencher-based assay for studying the ability of Dicer substrates to be processed by Dicer. By using a dsRNA probe labeled with Cy5-Iowa Black RQ, a panel of unlabeled test substrates can be rapidly assessed in heterologous competition assays without the need for electrophoresis or radiolabeling. This assay was piloted by evaluation of 196 unlabeled 27-mer Dicer substrates with various overhang structures in a purified Dicer enzyme assay system. Results indicate that Dicer has no preference for the sequence of RNA in the main double-stranded region of the substrate. However, a preference for Dicer substrate RNAs (D-siRNAs) containing purine/purine 3' overhang nucleotides was observed. These results demonstrate that the method is useful for studying the effects of modified nucleic acids in addition to rapidly accessing the influence of potential regulatory factors on Dicer processing.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Apr
pubmed:issn
1940-9818
pubmed:author
pubmed:issnType
Electronic
pubmed:volume
48
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
303-11
pubmed:dateRevised
2010-9-16
pubmed:meshHeading
pubmed:year
2010
pubmed:articleTitle
Continuous fluorescence-based method for assessing dicer cleavage efficiency reveals 3' overhang nucleotide preference.
pubmed:affiliation
Pfizer Research Technology Center, Cambridge, MA, USA. jonathan.dinitto@biogenidec.com
pubmed:publicationType
Journal Article