Targeted mutagenesis of the murine IRP1 and IRP2 genes reveals context-dependent RNA processing differences in vivo

被引：43

作者：

Galy, B ^{[1
]}

Ferring, D ^{[1
]}

Benesova, M ^{[1
]}

Benes, V ^{[1
]}

Hentze, MW ^{[1
]}

机构：

[1] European Mol Biol Lab, D-69117 Heidelberg, Germany

来源：

RNA | 2004年 / 10卷 / 07期

关键词：

alternative splicing; mRNA processing; gene trap; iron metabolism; NMD;

D O I：

10.1261/rna.7220704

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

We report the targeted mutagenesis of the murine iron regulatory protein (IRP)-1 and IRP2 genes, respectively, with a classical gene trap construct. Insertion of the targeting cassette into the second intron of either gene by homologous recombination interrupts their open reading frames near the N termini. Mice that are homozygous for the correctly modified IRP1 or IRP2 alleles, respectively, display a strong reduction (90%, IRP1(-/-)) or nondetectable levels (IRP2(-/-)) of the targeted proteins. Interestingly, the pre-mRNAs transcribed from the identical targeting cassettes are processed differently within the two different contexts. Detailed analysis of the respective products identifies the choice of alternative splice and 3' end processing sites in the same tissues in vivo. We discuss the implications for the understanding of RNA processing and for targeting strategies for functional genomics in the mouse.

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收藏

页码：1019 / 1025

页数：7

相关论文

共 27 条

[1] Efficient gene trap screening for novel developmental genes using IRESβgeo vector and in vitro preselection [J].

Bonaldo, P ;

Chowdhury, K ;

Stoykova, A ;

Torres, M ;

Gruss, P .

EXPERIMENTAL CELL RESEARCH, 1998, 244 (01) :125-136

[2] Iron regulatory proteins in pathobiology [J].

Cairo, G ;

Pietrangelo, A .

BIOCHEMICAL JOURNAL, 2000, 352 :241-250

[3] Evidence for the stochastic integration of gene trap vectors into the mouse germline [J].

Chowdhury, K ;

Bonaldo, P ;

Torres, M ;

Stoykova, A ;

Gruss, P .

NUCLEIC ACIDS RESEARCH, 1997, 25 (08) :1531-1536

[4] Abnormally spliced β-globin mRNAs:: a single point mutation generates transcripts sensitive and insensitive to nonsense-mediated mRNA decay [J].

Danckwardt, S ;

Neu-Yilik, G ;

Thermann, R ;

Frede, U ;

Hentze, MW ;

Kulozik, AE .

BLOOD, 2002, 99 (05) :1811-1816

[5] Flp recombinase promotes site-specific DNA recombination in embryonic stem cells and transgenic mice [J].

Dymecki, SM .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1996, 93 (12) :6191-6196

[6] Perinatal lethality of microtubule-associated protein 1B-deficient mice expressing alternative isoforms of the protein at low levels [J].

González-Billault, C ;

Demandt, E ;

Wandosell, F ;

Torres, M ;

Bonaldo, P ;

Stoykova, A ;

Chowdhury, K ;

Gruss, P ;

Avila, J ;

Sánchez, MP .

MOLECULAR AND CELLULAR NEUROSCIENCE, 2000, 16 (04) :408-421

[7] Mice with targeted disruption of the fatty acid transport protein 4 (Fatp 4, SlC27a4) gene show features of lethal restrictive dermopathy [J].

Herrmann, T ;

van der Hoeven, F ;

Gröne, HJ ;

Stewart, AF ;

Langbein, L ;

Kaiser, I ;

Liebisch, G ;

Gosch, I ;

Buchkremer, F ;

Drobnik, W ;

Schmitz, G ;

Stremmel, W .

JOURNAL OF CELL BIOLOGY, 2003, 161 (06) :1105-1115

[8] Targeted deletion of the gene encoding iron regulatory protein-2 causes misregulation of iron metabolism and neurodegenerative disease in mice [J].

LaVaute, T ;

Smith, S ;

Cooperman, S ;

Iwai, K ;

Land, W ;

Meyron-Holtz, E ;

Drake, SK ;

Miller, G ;

Abu-Asab, M ;

Tsokos, M ;

Switzer, R ;

Grinberg, A ;

Love, P ;

Tresser, N ;

Rouault, TA .

NATURE GENETICS, 2001, 27 (02) :209-214

[9] Genetic ablations of iron regulatory proteins 1 and 2 reveal why iron regulatory protein 2 dominates iron homeostasis [J].

Meyron-Holtz, EG ;

Ghosh, MC ;

Iwai, K ;

LaVaute, T ;

Brazzolotto, X ;

Berger, UV ;

Land, W ;

Ollivierre-Wilson, H ;

Grinberg, A ;

Love, P ;

Rouault, TA .

EMBO JOURNAL, 2004, 23 (02) :386-395

[10]

Nagy A, 2000, GENESIS, V26, P99, DOI 10.1002/(SICI)1526-968X(200002)26:2<99::AID-GENE1>3.0.CO