title: A Novel Method for Gene-Specific Enhancement of Protein Translation by Targeting 5'UTRs of Selected Tumor Suppressors.
abstract: Translational control is a mechanism of protein synthesis regulation emerging as an important target for new therapeutics. Naturally occurring microRNAs and synthetic small inhibitory RNAs (siRNAs) are the most recognized regulatory molecules acting via RNA interference. Surprisingly, recent studies have shown that interfering RNAs may also activate gene transcription via the newly discovered phenomenon of small RNA-induced gene activation (RNAa). Thus far, the small activating RNAs (saRNAs) have only been demonstrated as promoter-specific transcriptional activators.
  
  
  
  We demonstrate that oligonucleotide-based trans-acting factors can also specifically enhance gene expression at the level of protein translation by acting at sequence-specific targets within the messenger RNA 5'-untranslated region (5'UTR). We designed a set of short synthetic oligonucleotides (dGoligos), specifically targeting alternatively spliced 5'UTRs in transcripts expressed from the THRB and CDKN2A suppressor genes. The in vitro translation efficiency of reporter constructs containing alternative TRβ1 5'UTRs was increased by up to more than 55-fold following exposure to specific dGoligos. Moreover, we found that the most folded 5'UTR has higher translational regulatory potential when compared to the weakly folded TRβ1 variant. This suggests such a strategy may be especially applied to enhance translation from relatively inactive transcripts containing long 5'UTRs of complex structure.
  
  
  
  This report represents the first method for gene-specific translation enhancement using selective trans-acting factors designed to target specific 5'UTR cis-acting elements. This simple strategy may be developed further to complement other available methods for gene expression regulation including gene silencing. The dGoligo-mediated translation-enhancing approach has the potential to be transferred to increase the translation efficiency of any suitable target gene and may have future application in gene therapy strategies to enhance expression of proteins including tumor suppressors.
authors.affiliation: The Centre of Postgraduate Medical Education, Department of Biochemistry and Molecular Biology, ul. Marymoncka 99/103, 01-813, Warsaw, Poland.|BioTe21, Laboratory of Molecular Medical Biology, ul. Krolowej Jadwigi 33/3b, 30-209, Cracow, Poland.
authors.forename: Adam
authors.initials: A
authors.lastname: Master
authors.affiliation: The Centre of Postgraduate Medical Education, Department of Biochemistry and Molecular Biology, ul. Marymoncka 99/103, 01-813, Warsaw, Poland.|Centre of New Technologies, University of Warsaw, Banacha 2c, 02-089, Warsaw, Poland.|Genomic Medicine, Medical University of Warsaw, Zwirki i Wigury 61, 02-091, Warsaw, Poland.
authors.forename: Anna
authors.initials: A
authors.lastname: Wójcicka
authors.affiliation: BioTe21, Laboratory of Molecular Medical Biology, ul. Krolowej Jadwigi 33/3b, 30-209, Cracow, Poland.
authors.forename: Kamilla
authors.initials: K
authors.lastname: Giżewska
authors.affiliation: The Centre of Postgraduate Medical Education, Department of Biochemistry and Molecular Biology, ul. Marymoncka 99/103, 01-813, Warsaw, Poland.
authors.forename: Piotr
authors.initials: P
authors.lastname: Popławski
authors.affiliation: Molecular Endocrinology Group, Department of Medicine, Imperial College London, Hammersmith Campus, London, W12 0NN, United Kingdom.
authors.forename: Graham R
authors.initials: GR
authors.lastname: Williams
authors.affiliation: The Centre of Postgraduate Medical Education, Department of Biochemistry and Molecular Biology, ul. Marymoncka 99/103, 01-813, Warsaw, Poland.|Centre of New Technologies, University of Warsaw, Banacha 2c, 02-089, Warsaw, Poland.
authors.forename: Alicja
authors.initials: A
authors.lastname: Nauman
mesh_terms: D020121:5' Untranslated Regions / Q000235:genetics*; D001483:Base Sequence; D045744:Cell Line, Tumor; D019941:Cyclin-Dependent Kinase Inhibitor p16 / Q000378:metabolism; D005786:Gene Expression Regulation; D016147:Genes, Tumor Suppressor; D005821:Genetic Techniques; D006801:Humans; D009690:Nucleic Acid Conformation; D014176:Protein Biosynthesis / Q000235:genetics*; D011988:Receptors, Thyroid Hormone / Q000235:genetics; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics; D013816:Thermodynamics
chemical_list: D020121:5' Untranslated Regions; D019941:Cyclin-Dependent Kinase Inhibitor p16; D011988:Receptors, Thyroid Hormone