title: Targeted 'knockdown' of spliceosome function in mammalian cells.
abstract: The existence of two sophisticated parallel splicing machineries in multicellular organisms has raised intriguing questions--ranging from their impact on proteome expansion to the evolution of splicing and of metazoan genomes. Exploring roles for the distinct splicing systems in vivo has, however, been restricted by the lack of techniques to selectively inhibit their function in cells. In this study, we show that morpholino oligomers complementary to the branch-site recognition elements of U2 or U12 small nuclear RNA specifically suppress the function of the two splicing systems in mammalian cells. The data provide the first evidence for a role of distinct spliceosomes in pre-mRNA splicing from endogenous mammalian genes and establish a tool to define roles for the different splicing machineries in vivo.
authors.affiliation: Forschungszentrum Karlsruhe GmbH, Institut für Toxikologie und Genetik Postfach 3640, D-76021 Karlsruhe, Germany.
authors.forename: Nathalie
authors.initials: N
authors.lastname: Matter
authors.forename: Harald
authors.initials: H
authors.lastname: König
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D006801:Humans; D019169:Jurkat Cells; D051379:Mice; D020320:Oligoribonucleotides, Antisense / Q000494:pharmacology*; D012326:RNA Splicing / Q000187:drug effects; D012342:RNA, Small Nuclear / Q000037:antagonists & inhibitors* / Q000502:physiology; D017381:Spliceosomes / Q000187:drug effects / Q000502:physiology*
chemical_list: D020320:Oligoribonucleotides, Antisense; D012342:RNA, Small Nuclear; C096986:U12 small nuclear RNA; C405484:U2 small nuclear RNA
references.citation: Biochim Biophys Acta. 1999 Dec 10;1489(1):141-58
references.pmid: 10807004
references.citation: Genes Dev. 1999 Apr 1;13(7):851-63
references.pmid: 10197985
references.citation: Nat Genet. 2000 Oct;26(2):216-20
references.pmid: 11017081
references.citation: Nucleic Acids Res. 2001 Oct 1;29(19):4006-13
references.pmid: 11574683
references.citation: Mol Cell. 2002 Feb;9(2):439-46
references.pmid: 11864616
references.citation: Dev Biol. 2002 Mar 15;243(2):209-14
references.pmid: 11884031
references.citation: Nat Biotechnol. 2002 Apr;20(4):353-8
references.pmid: 11923840
references.citation: Mech Dev. 2002 Oct;118(1-2):57-63
references.pmid: 12351170
references.citation: Nat Biotechnol. 2002 Dec;20(12):1228-33
references.pmid: 12426578
references.citation: Nature. 2002 Dec 12;420(6916):691-5
references.pmid: 12478298
references.citation: Dev Dyn. 2003 Mar;226(3):470-7
references.pmid: 12619133
references.citation: Mol Cell. 2003 Jul;12(1):5-14
references.pmid: 12887888
references.citation: Nat Rev Mol Cell Biol. 2003 Dec;4(12):960-70
references.pmid: 14685174
references.citation: Science. 2003 Dec 19;302(5653):2141-4
references.pmid: 14684825
references.citation: RNA. 2004 Aug;10(8):1243-50
references.pmid: 15272120
references.citation: Proc Natl Acad Sci U S A. 1988 Dec;85(23):8885-9
references.pmid: 2973606
references.citation: Cell. 1989 Jul 28;58(2):383-90
references.pmid: 2526684
references.citation: Mol Cell Biol. 1990 Jan;10(1):84-94
references.pmid: 2136768
references.citation: Nucleic Acids Res. 1990 May 25;18(10):3015-9
references.pmid: 2349097
references.citation: Genes Dev. 1993 Oct;7(10):1926-36
references.pmid: 8405999
references.citation: Science. 1996 Mar 22;271(5256):1716-8
references.pmid: 8596930
references.citation: Science. 1996 Sep 20;273(5282):1706-9
references.pmid: 8781232
references.citation: RNA. 1997 Mar;3(3):227-33
references.pmid: 9056760
references.citation: Cell. 1997 Dec 26;91(7):875-9
references.pmid: 9428511
references.citation: Mol Cell. 1997 Dec;1(1):151-60
references.pmid: 9659912
references.citation: Mol Cell. 1998 Dec;2(6):773-85
references.pmid: 9885565
references.citation: Proc Natl Acad Sci U S A. 2000 Aug 15;97(17):9591-6
references.pmid: 10944225