Click here to close
Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly.
We suggest using a current version of Chrome,
FireFox, or Safari.
Proc Natl Acad Sci U S A
2013 Nov 05;11045:18285-90. doi: 10.1073/pnas.1306243110.
Show Gene links
Show Anatomy links
Correction of mutations within the cystic fibrosis transmembrane conductance regulator by site-directed RNA editing.
Montiel-Gonzalez MF, Vallecillo-Viejo I, Yudowski GA, Rosenthal JJ.
???displayArticle.abstract???
Adenosine deaminases that act on RNA are a conserved family of enzymes that catalyze a natural process of site-directed mutagenesis. Biochemically, they convert adenosine to inosine, a nucleotide that is read as guanosine during translation; thus when editing occurs in mRNAs, codons can be recoded and the changes can alter protein function. By removing the endogenous targeting domains from human adenosine deaminase that acts on RNA 2 and replacing them with an antisense RNA oligonucleotide, we have engineered a recombinant enzyme that can be directed to edit anywhere along the RNA registry. Here we demonstrate that this enzyme can efficiently and selectively edit a single adenosine. As proof of principle in vitro, we correct a premature termination codon in mRNAs encoding the cystic fibrosis transmembrane conductance regulator anion channel. In Xenopus oocytes, we show that a genetically encoded version of our editase can correct cystic fibrosis transmembrane conductance regulator mRNA, restore full-length protein, and reestablish functional chloride currents across the plasma membrane. Finally, in a human cell line, we show that a genetically encoded version of our editase and guide RNA can correct a nonfunctional version of enhanced green fluorescent protein, which contains a premature termination codon. This technology should spearhead powerful approaches to correcting a wide variety of genetic mutations and fine-tuning protein function through targeted nucleotide deamination.
Austin,
Designed arginine-rich RNA-binding peptides with picomolar affinity.
2002, Pubmed
Austin,
Designed arginine-rich RNA-binding peptides with picomolar affinity.
2002,
Pubmed Balassopoulou,
A novel nonsense mutation identified in the first nucleotide binding fold of the CFTR gene in a Greek patient.
1994,
Pubmed BASILIO,
Synthetic polynucleotides and the amino acid code. V.
1962,
Pubmed Bass,
A developmentally regulated activity that unwinds RNA duplexes.
1987,
Pubmed
,
Xenbase Bass,
An unwinding activity that covalently modifies its double-stranded RNA substrate.
1988,
Pubmed
,
Xenbase Bear,
Cl- channel activity in Xenopus oocytes expressing the cystic fibrosis gene.
1991,
Pubmed
,
Xenbase Bhalla,
Control of human potassium channel inactivation by editing of a small mRNA hairpin.
2004,
Pubmed Burns,
Regulation of serotonin-2C receptor G-protein coupling by RNA editing.
1997,
Pubmed Chattopadhyay,
Bipartite function of a small RNA hairpin in transcription antitermination in bacteriophage lambda.
1995,
Pubmed Cheng,
Defective intracellular transport and processing of CFTR is the molecular basis of most cystic fibrosis.
1990,
Pubmed Cilley,
Analysis of bacteriophage N protein and peptide binding to boxB RNA using polyacrylamide gel coelectrophoresis (PACE).
1997,
Pubmed Colina,
Regulation of Na+/K+ ATPase transport velocity by RNA editing.
2010,
Pubmed Egan,
Defective regulation of outwardly rectifying Cl- channels by protein kinase A corrected by insertion of CFTR.
1992,
Pubmed Eggington,
Predicting sites of ADAR editing in double-stranded RNA.
2011,
Pubmed Gehring,
Exon-junction complex components specify distinct routes of nonsense-mediated mRNA decay with differential cofactor requirements.
2005,
Pubmed Gregory,
Expression and characterization of the cystic fibrosis transmembrane conductance regulator.
1990,
Pubmed Gribble,
A novel method for measurement of submembrane ATP concentration.
2000,
Pubmed
,
Xenbase Herb,
Q/R site editing in kainate receptor GluR5 and GluR6 pre-mRNAs requires distant intronic sequences.
1996,
Pubmed Higuchi,
RNA editing of AMPA receptor subunit GluR-B: a base-paired intron-exon structure determines position and efficiency.
1993,
Pubmed Karijolich,
Converting nonsense codons into sense codons by targeted pseudouridylation.
2011,
Pubmed Keegan,
Purification and assay of ADAR activity.
2007,
Pubmed Kim,
Molecular cloning of cDNA for double-stranded RNA adenosine deaminase, a candidate enzyme for nuclear RNA editing.
1994,
Pubmed Lehmann,
Double-stranded RNA adenosine deaminases ADAR1 and ADAR2 have overlapping specificities.
2000,
Pubmed
,
Xenbase Macbeth,
Inositol hexakisphosphate is bound in the ADAR2 core and required for RNA editing.
2005,
Pubmed Melcher,
A mammalian RNA editing enzyme.
1996,
Pubmed Nishikura,
Functions and regulation of RNA editing by ADAR deaminases.
2010,
Pubmed O'Connell,
Purification of human double-stranded RNA-specific editase 1 (hRED1) involved in editing of brain glutamate receptor B pre-mRNA.
1997,
Pubmed O'Connell,
Purification of native and recombinant double-stranded RNA-specific adenosine deaminases.
1998,
Pubmed Palavicini,
An extra double-stranded RNA binding domain confers high activity to a squid RNA editing enzyme.
2009,
Pubmed Peters,
The behavioral phenotype in MECP2 duplication syndrome: a comparison with idiopathic autism.
2013,
Pubmed Rinkevich,
Antisense sequencing improves the accuracy and precision of A-to-I editing measurements using the peak height ratio method.
2012,
Pubmed Rosenthal,
A-to-I RNA editing: effects on proteins key to neural excitability.
2012,
Pubmed Schwarz,
Cystic fibrosis mutation analysis: report from 22 U.K. regional genetics laboratories.
1995,
Pubmed Sommer,
RNA editing in brain controls a determinant of ion flow in glutamate-gated channels.
1991,
Pubmed Stafforst,
An RNA-deaminase conjugate selectively repairs point mutations.
2012,
Pubmed Tan,
Structural variety of arginine-rich RNA-binding peptides.
1995,
Pubmed Venglarik,
ATP alters current fluctuations of cystic fibrosis transmembrane conductance regulator: evidence for a three-state activation mechanism.
1994,
Pubmed Woolf,
Toward the therapeutic editing of mutated RNA sequences.
1995,
Pubmed
,
Xenbase
