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Blood, 18 June 2009, Vol. 113, No. 25, pp. 6461-6464. Prepublished online as a Blood First Edition Paper on April 22, 2009; DOI 10.1182/blood-2009-03-207613. This Article Full Text Full Text (PDF) All Versions of this Article: blood-2009-03-207613v1 113/25/6461    most recent Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Rights and Permissions Citing Articles Citing Articles via CrossRef Google Scholar Articles by Pinotti, M. Articles by Bernardi, F. PubMed PubMed Citation Articles by Pinotti, M. Articles by Bernardi, F. Related Collections Thrombosis and Hemostasis Brief Reports Social Bookmarking                   What's this?  Previous Article  |  Table of Contents  |  Next Article  THROMBOSIS AND HEMOSTASIS Brief report Rescue of coagulation factor VII function by the U1+5A snRNA Mirko Pinotti1, Dario Balestra1, Lara Rizzotto1, Iva Maestri2, Franco Pagani3, and Francesco Bernardi1 Departments of 1 Biochemistry and Molecular Biology and 2 Experimental and Diagnostic Medicine, University of Ferrara, Ferrara; and 3 International Centre for Genetic Engineering and Biotechnology, Trieste, Italy Our previous studies with genomic minigenes have demonstrated that an engineered small nuclear RNA-U1 (U1+5a) partially rescued coagulation factor VII (FVII) mRNA processing impaired by the 9726+5G>A mutation. Here, to evaluate the U1+5a effects on FVII function, we devised a full-length FVII splicing-competent construct (pSCFVII-wt). This construct drove in COS-1 cells the synthesis of properly processed FVII transcripts and of secreted functional FVII (23 ± 4 ng/mL), which were virtually undetectable upon introduction of the 9726+5G>A mutation (pSCFVII-9726+5a). Cotransfection of pSCFVII-9726+5a with pU1+5a resulted in a partial rescue of FVII splicing and protein biosynthesis. The level increase in medium was dose dependent and, with a molar excess (1.5x) of pU1+5a, reached 9.5% plus or minus 3.2% (5.0 ± 2.8 ng/mL) of FVII-wt coagulant activity. These data provide the first insights into the U1-snRNA–mediated rescue of donor splice sites at protein level, thus further highlighting its therapeutic implications in bleeding disorders, which would benefit even from tiny increase of functional levels. CiteULike    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

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