Literature DB >> 21726522

Fluorescence-based evaluation of shRNA efficacy.

B J Naughton1, Dawn D Han, Howard H Gu.   

Abstract

RNA interference is a cellular mechanism regulating levels of mRNAs. It has been widely exploited to knock down specific protein targets. The selected interfering RNA sequence greatly influences its ability to knock down the target. Here we present a method for constructing multiple testing plasmids which express small hairpin RNAs (shRNA) targeting different regions of an mRNA. A simple fluorescence test in cultured cells allows convenient evaluation of mRNA knockdown by many different shRNAs on 96-well plates. We show that software predicted shRNAs have varying efficacies and only 2 of the 7 tested shRNAs significantly knocked down their targets.
Copyright © 2011 Elsevier Inc. All rights reserved.

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Year:  2011        PMID: 21726522      PMCID: PMC3143307          DOI: 10.1016/j.ab.2011.06.008

Source DB:  PubMed          Journal:  Anal Biochem        ISSN: 0003-2697            Impact factor:   3.365


  7 in total

1.  Stable suppression of gene expression by RNAi in mammalian cells.

Authors:  Patrick J Paddison; Amy A Caudy; Gregory J Hannon
Journal:  Proc Natl Acad Sci U S A       Date:  2002-01-29       Impact factor: 11.205

2.  Short hairpin RNAs (shRNAs) induce sequence-specific silencing in mammalian cells.

Authors:  Patrick J Paddison; Amy A Caudy; Emily Bernstein; Gregory J Hannon; Douglas S Conklin
Journal:  Genes Dev       Date:  2002-04-15       Impact factor: 11.361

3.  Bright monomeric red fluorescent protein with an extended fluorescence lifetime.

Authors:  Ekaterina M Merzlyak; Joachim Goedhart; Dmitry Shcherbo; Mariya E Bulina; Aleksandr S Shcheglov; Arkady F Fradkov; Anna Gaintzeva; Konstantin A Lukyanov; Sergey Lukyanov; Theodorus W J Gadella; Dmitriy M Chudakov
Journal:  Nat Methods       Date:  2007-06-17       Impact factor: 28.547

Review 4.  Control of translation and mRNA degradation by miRNAs and siRNAs.

Authors:  Marco Antonio Valencia-Sanchez; Jidong Liu; Gregory J Hannon; Roy Parker
Journal:  Genes Dev       Date:  2006-03-01       Impact factor: 11.361

5.  New unstable variants of green fluorescent protein for studies of transient gene expression in bacteria.

Authors:  J B Andersen; C Sternberg; L K Poulsen; S P Bjorn; M Givskov; S Molin
Journal:  Appl Environ Microbiol       Date:  1998-06       Impact factor: 4.792

6.  Use of bicistronic vectors in combination with flow cytometry to screen for effective small interfering RNA target sequences.

Authors:  Naoka Kamio; Hideyo Hirai; Eishi Ashihara; Daniel G Tenen; Taira Maekawa; Jiro Imanishi
Journal:  Biochem Biophys Res Commun       Date:  2010-02-10       Impact factor: 3.575

7.  A resource for large-scale RNA-interference-based screens in mammals.

Authors:  Patrick J Paddison; Jose M Silva; Douglas S Conklin; Mike Schlabach; Mamie Li; Shola Aruleba; Vivekanand Balija; Andy O'Shaughnessy; Lidia Gnoj; Kim Scobie; Kenneth Chang; Thomas Westbrook; Michele Cleary; Ravi Sachidanandam; W Richard McCombie; Stephen J Elledge; Gregory J Hannon
Journal:  Nature       Date:  2004-03-25       Impact factor: 49.962

  7 in total
  3 in total

1.  Specific knockdown of the D2 long dopamine receptor variant.

Authors:  Bart J Naughton; Keerthi Thirtamara-Rajamani; Chuansong Wang; Matthew J During; Howard H Gu
Journal:  Neuroreport       Date:  2012-01-04       Impact factor: 1.837

2.  Engineering human megakaryocytic microparticles for targeted delivery of nucleic acids to hematopoietic stem and progenitor cells.

Authors:  Chen-Yuan Kao; Eleftherios T Papoutsakis
Journal:  Sci Adv       Date:  2018-11-07       Impact factor: 14.136

Review 3.  Recombinant Adeno-Associated Viral Vectors (rAAV)-Vector Elements in Ocular Gene Therapy Clinical Trials and Transgene Expression and Bioactivity Assays.

Authors:  Thilo M Buck; Jan Wijnholds
Journal:  Int J Mol Sci       Date:  2020-06-12       Impact factor: 5.923

  3 in total

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