Source:http://linkedlifedata.com/resource/pubmed/id/11127576
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| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
22
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| pubmed:dateCreated |
2000-12-20
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| pubmed:abstractText |
Most messenger RNA precursors (pre-mRNA) undergo cis-splicing in which introns are excised and the adjoining exons from a single pre-mRNA are ligated together to form mature messenger RNA. This reaction is driven by a complex known as the spliceosome. Spliceosomes can also combine sequences from two independently transcribed pre-mRNAs in a process known as trans-splicing. Spliceosome-mediated RNA trans-splicing (SMaRT) is an emerging technology in which RNA pre-therapeutic molecules (PTMs) are designed to recode a specific pre-mRNA by suppressing cis-splicing while enhancing trans-splicing between the PTM and its pre-mRNA target. This study examined the feasibility of SMaRT as a potential therapy for genetic diseases to correct mutations using cystic fibrosis (CF) as an example. We used several versions of a cystic fibrosis transmembrane conductance regulator (CFTR) mini-gene expressing mutant (deltaF508) pre-mRNA targets and tested this against a number of PTMs capable of binding to the CFTR target intron 9 and trans-splicing in the normal coding sequences for exons 10-24 (containing F508). When 293T cells were cotransfected with both constructs, they produced a trans-spliced mRNA in which normal exon 10-24 replaced mutant exon 10. To test whether SMaRT produced mature CFTR protein, proteins were immunoprecipitated from lysates of cotransfected cells and detected by Western blotting and PKA-phosphorylation. Tryptic phosphopeptide mapping confirmed the identity of CFTR. This proof-of-concept study demonstrates that exon replacement by SMaRT can repair an abnormal pre-mRNA associated with a genetic disease.
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| pubmed:grant | |
| pubmed:language |
eng
|
| pubmed:journal | |
| pubmed:citationSubset |
IM
|
| pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/CFTR protein, human,
http://linkedlifedata.com/resource/pubmed/chemical/Cation Exchange Resins,
http://linkedlifedata.com/resource/pubmed/chemical/Cystic Fibrosis Transmembrane...,
http://linkedlifedata.com/resource/pubmed/chemical/Lipids,
http://linkedlifedata.com/resource/pubmed/chemical/Lipofectamine,
http://linkedlifedata.com/resource/pubmed/chemical/RNA Precursors,
http://linkedlifedata.com/resource/pubmed/chemical/RNA Splice Sites
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| pubmed:status |
MEDLINE
|
| pubmed:month |
Nov
|
| pubmed:issn |
0969-7128
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| pubmed:author | |
| pubmed:issnType |
Print
|
| pubmed:volume |
7
|
| pubmed:owner |
NLM
|
| pubmed:authorsComplete |
Y
|
| pubmed:pagination |
1885-95
|
| pubmed:dateRevised |
2007-11-14
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| pubmed:meshHeading |
pubmed-meshheading:11127576-Blotting, Western,
pubmed-meshheading:11127576-Cation Exchange Resins,
pubmed-meshheading:11127576-Cell Line,
pubmed-meshheading:11127576-Colon,
pubmed-meshheading:11127576-Cystic Fibrosis,
pubmed-meshheading:11127576-Cystic Fibrosis Transmembrane Conductance Regulator,
pubmed-meshheading:11127576-Exons,
pubmed-meshheading:11127576-Feasibility Studies,
pubmed-meshheading:11127576-Gene Therapy,
pubmed-meshheading:11127576-Genetic Engineering,
pubmed-meshheading:11127576-Humans,
pubmed-meshheading:11127576-Kidney,
pubmed-meshheading:11127576-Lipids,
pubmed-meshheading:11127576-RNA Precursors,
pubmed-meshheading:11127576-RNA Splice Sites,
pubmed-meshheading:11127576-Reverse Transcriptase Polymerase Chain Reaction,
pubmed-meshheading:11127576-Spliceosomes,
pubmed-meshheading:11127576-Transfection
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| pubmed:year |
2000
|
| pubmed:articleTitle |
Repair of CFTR mRNA by spliceosome-mediated RNA trans-splicing.
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| pubmed:affiliation |
Intronn, LLC, Durham, NC 27701, USA.
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| pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.,
Research Support, Non-U.S. Gov't
|