Literature DB >> 18802435

Design and delivery of antisense oligonucleotides to block microRNA function in cultured Drosophila and human cells.

Michael D Horwich1, Phillip D Zamore.   

Abstract

MicroRNAs (miRNAs), approximately 22-nt RNAs that mediate post-transcriptional regulation of mRNAs in animals and plants, are a diverse class of regulatory genes whose specific biological functions are largely unknown. Here we detail a protocol to design and introduce into cultured Drosophila and human cells sequence-specific antisense oligonucleotides (ASOs) that block the function of individual miRNAs. Coupled with recent studies that catalog the miRNAs expressed in diverse cultured cells, our method offers a rapid (<1 week) approach to validate miRNA targets and to study the cellular functions of individual human and Drosophila miRNAs. ASO-based inactivation of miRNAs is faster and simpler than comparable genetic or 'sponge'-based approaches, for which extensive recombinant DNA manipulation is required. We present our ASO design principles and an optimized transfection protocol in which transfection efficiency of Drosophila Schneider 2 cells can approach 100%. Our 3'-cholesterol-modified ASOs have enhanced potency, allowing miRNA inhibition for at least 7 d from a single transfection.

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Year:  2008        PMID: 18802435      PMCID: PMC2559958          DOI: 10.1038/nprot.2008.145

Source DB:  PubMed          Journal:  Nat Protoc        ISSN: 1750-2799            Impact factor:   13.491


  104 in total

1.  A cellular function for the RNA-interference enzyme Dicer in the maturation of the let-7 small temporal RNA.

Authors:  G Hutvágner; J McLachlan; A E Pasquinelli; E Bálint; T Tuschl; P D Zamore
Journal:  Science       Date:  2001-07-12       Impact factor: 47.728

2.  The small RNA profile during Drosophila melanogaster development.

Authors:  Alexei A Aravin; Mariana Lagos-Quintana; Abdullah Yalcin; Mihaela Zavolan; Debora Marks; Ben Snyder; Terry Gaasterland; Jutta Meyer; Thomas Tuschl
Journal:  Dev Cell       Date:  2003-08       Impact factor: 12.270

3.  RNAi-mediated allelic trans-interaction at the imprinted Rtl1/Peg11 locus.

Authors:  Erica Davis; Florian Caiment; Xavier Tordoir; Jérôme Cavaillé; Anne Ferguson-Smith; Noelle Cockett; Michel Georges; Carole Charlier
Journal:  Curr Biol       Date:  2005-04-26       Impact factor: 10.834

4.  A genetic screen implicates miRNA-372 and miRNA-373 as oncogenes in testicular germ cell tumors.

Authors:  P Mathijs Voorhoeve; Carlos le Sage; Mariette Schrier; Ad J M Gillis; Hans Stoop; Remco Nagel; Ying-Poi Liu; Josyanne van Duijse; Jarno Drost; Alexander Griekspoor; Eitan Zlotorynski; Norikazu Yabuta; Gabriella De Vita; Hiroshi Nojima; Leendert H J Looijenga; Reuven Agami
Journal:  Cell       Date:  2006-03-24       Impact factor: 41.582

5.  The tumor suppressor microRNA let-7 represses the HMGA2 oncogene.

Authors:  Yong Sun Lee; Anindya Dutta
Journal:  Genes Dev       Date:  2007-04-16       Impact factor: 11.361

6.  Therapeutic silencing of an endogenous gene by systemic administration of modified siRNAs.

Authors:  Jürgen Soutschek; Akin Akinc; Birgit Bramlage; Klaus Charisse; Rainer Constien; Mary Donoghue; Sayda Elbashir; Anke Geick; Philipp Hadwiger; Jens Harborth; Matthias John; Venkitasamy Kesavan; Gary Lavine; Rajendra K Pandey; Timothy Racie; Kallanthottathil G Rajeev; Ingo Röhl; Ivanka Toudjarska; Gang Wang; Silvio Wuschko; David Bumcrot; Victor Koteliansky; Stefan Limmer; Muthiah Manoharan; Hans-Peter Vornlocher
Journal:  Nature       Date:  2004-11-11       Impact factor: 49.962

7.  Interferon modulation of cellular microRNAs as an antiviral mechanism.

Authors:  Irene M Pedersen; Guofeng Cheng; Stefan Wieland; Stefano Volinia; Carlo M Croce; Francis V Chisari; Michael David
Journal:  Nature       Date:  2007-10-18       Impact factor: 49.962

8.  A mammalian microRNA expression atlas based on small RNA library sequencing.

Authors:  Pablo Landgraf; Mirabela Rusu; Robert Sheridan; Alain Sewer; Nicola Iovino; Alexei Aravin; Sébastien Pfeffer; Amanda Rice; Alice O Kamphorst; Markus Landthaler; Carolina Lin; Nicholas D Socci; Leandro Hermida; Valerio Fulci; Sabina Chiaretti; Robin Foà; Julia Schliwka; Uta Fuchs; Astrid Novosel; Roman-Ulrich Müller; Bernhard Schermer; Ute Bissels; Jason Inman; Quang Phan; Minchen Chien; David B Weir; Ruchi Choksi; Gabriella De Vita; Daniela Frezzetti; Hans-Ingo Trompeter; Veit Hornung; Grace Teng; Gunther Hartmann; Miklos Palkovits; Roberto Di Lauro; Peter Wernet; Giuseppe Macino; Charles E Rogler; James W Nagle; Jingyue Ju; F Nina Papavasiliou; Thomas Benzing; Peter Lichter; Wayne Tam; Michael J Brownstein; Andreas Bosio; Arndt Borkhardt; James J Russo; Chris Sander; Mihaela Zavolan; Thomas Tuschl
Journal:  Cell       Date:  2007-06-29       Impact factor: 41.582

9.  microRNA target predictions across seven Drosophila species and comparison to mammalian targets.

Authors:  Dominic Grün; Yi-Lu Wang; David Langenberger; Kristin C Gunsalus; Nikolaus Rajewsky
Journal:  PLoS Comput Biol       Date:  2005-06-24       Impact factor: 4.475

10.  Targeted inhibition of miRNA maturation with morpholinos reveals a role for miR-375 in pancreatic islet development.

Authors:  Wigard P Kloosterman; Anne K Lagendijk; René F Ketting; Jon D Moulton; Ronald H A Plasterk
Journal:  PLoS Biol       Date:  2007-08       Impact factor: 8.029

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  43 in total

Review 1.  MicroRNAs and their roles in osteoclast differentiation.

Authors:  Zhuying Xia; Chao Chen; Peng Chen; Hui Xie; Xianghang Luo
Journal:  Front Med       Date:  2011-12-27       Impact factor: 4.592

2.  Use of target protector morpholinos to analyze the physiological roles of specific miRNA-mRNA pairs in vivo.

Authors:  Alison A Staton; Antonio J Giraldez
Journal:  Nat Protoc       Date:  2011-12-01       Impact factor: 13.491

Review 3.  Role of microRNAs in endothelial inflammation and senescence.

Authors:  Bing Qin; Huan Yang; Bo Xiao
Journal:  Mol Biol Rep       Date:  2011-09-28       Impact factor: 2.316

Review 4.  Exploiting and antagonizing microRNA regulation for therapeutic and experimental applications.

Authors:  Brian D Brown; Luigi Naldini
Journal:  Nat Rev Genet       Date:  2009-08       Impact factor: 53.242

Review 5.  Insect microRNAs: biogenesis, expression profiling and biological functions.

Authors:  Keira Lucas; Alexander S Raikhel
Journal:  Insect Biochem Mol Biol       Date:  2012-11-16       Impact factor: 4.714

Review 6.  Invoking the power of thrombospondins: regulation of thrombospondins expression.

Authors:  Olga Stenina-Adognravi
Journal:  Matrix Biol       Date:  2014-02-25       Impact factor: 11.583

Review 7.  MicroRNAs in ovarian carcinomas.

Authors:  Neetu Dahiya; Patrice J Morin
Journal:  Endocr Relat Cancer       Date:  2010-01-29       Impact factor: 5.678

Review 8.  Sequence-non-specific effects of RNA interference triggers and microRNA regulators.

Authors:  Marta Olejniczak; Paulina Galka; Wlodzimierz J Krzyzosiak
Journal:  Nucleic Acids Res       Date:  2009-10-20       Impact factor: 16.971

9.  Specificity and functionality of microRNA inhibitors.

Authors:  Barbara Robertson; Andrew B Dalby; Jon Karpilow; Anastasia Khvorova; Devin Leake; Annaleen Vermeulen
Journal:  Silence       Date:  2010-04-01

10.  microRNAs: a role in drug resistance in parasitic nematodes?

Authors:  Eileen Devaney; Alan D Winter; Collette Britton
Journal:  Trends Parasitol       Date:  2010-06-11
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