A comparison of methods for successful triggering of gene silencing in Coprinus cinereus
Heneghan, Mary N., Costa, Ana M. S. B., Challen, Michael P., Mills, P. R. (Peter R.), Bailey, Andy and Foster, Gary D.. (2007) A comparison of methods for successful triggering of gene silencing in Coprinus cinereus. Molecular Biotechnology, Vol.35 (No.3). pp. 283-294. ISSN 1073-6085Full text not available from this repository.
Official URL: http://dx.doi.org/10.1007/s12033-007-9008-5
Post-transcriptional gene-silencing methods (PTGS), including RNAi, are becoming pervasive in functional genomics. To advance analysis of the recently sequenced Coprinus cinereus genome, a high throughput gene silencing method is essential. We have exploited the GFP reporter gene to evaluate and quantify efficacy of three different silencing strategies. Modular constructs that encompassed antisense, untranslatable sense, and RNAi-mediating hairpin sequences, were transformed into a GFP-expressing host strain. Transformants exhibiting strong downregulation and partial suppression of GFP were recovered with all three constructs. Analyses of protein and transcriptional nucleic acids revealed that the antosense and hairpin sequences yielded similar levels of GFP suppression, and were both more efficient than untranslatable sense sequence. Our antisense vectors will expedite functional characterisation of C. cinereus and the modular nature of the constructs should permit exploitation of directional cDNA libraries for high throughput screening.
|Item Type:||Journal Article|
|Subjects:||Q Science > QD Chemistry
T Technology > TP Chemical technology
|Divisions:||Faculty of Science > Life Sciences (2010- ) > Warwick HRI (2004-2010)|
|Journal or Publication Title:||Molecular Biotechnology|
|Publisher:||Humana Press, Inc.|
|Number of Pages:||14|
|Page Range:||pp. 283-294|
|Access rights to Published version:||Restricted or Subscription Access|
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