Utrophin up-regulation by an artificial transcription factor in transgenic mice.
Duchenne Muscular Dystrophy (DMD) is a severe muscle degenerative disease, due to absence of dystrophin. There is currently no effective treatment for DMD. Our aim is to up-regulate the expression level of the dystrophin related gene utrophin in DMD, complementing in this way the lack of dystrophin functions. To this end we designed and engineered several synthetic zinc finger based transcription factors. In particular, we have previously shown that the artificial three zinc finger protein named Jazz, fused with the appropriate effector domain, is able to drive the transcription of a test gene from the utrophin promoter "A". Here we report on the characterization of Vp16-Jazz-transgenic mice that specifically over-express the utrophin gene at the muscular level. A Chromatin Immunoprecipitation assay (ChIP) demonstrated the effective access/binding of the Jazz protein to active chromatin in mouse muscle and Vp16-Jazz was shown to be able to up-regulate endogenous utrophin gene expression by immunohistochemistry, western blot analyses and real-time PCR. To our knowledge, this is the first example of a transgenic mouse expressing an artificial gene coding for a zinc finger based transcription factor. The achievement of Vp16-Jazz transgenic mice validates the strategy of transcriptional targeting of endogenous genes and could represent an exclusive animal model for use in drug discovery and therapeutics.
Mattei
Elisabetta
E
Istituto di Neurobiologia e Medicina Molecolare, Consiglio Nazionale delle Ricerche, European Brain Research Institute, Rome, Italy.
Corbi
Nicoletta
N
Di Certo
Maria Grazia
MG
Strimpakos
Georgios
G
Severini
Cinzia
C
Onori
Annalisa
A
Desantis
Agata
A
Libri
Valentina
V
Buontempo
Serena
S
Floridi
Aristide
A
Fanciulli
Maurizio
M
Baban
Dilair
D
Davies
Kay E
KE
Passananti
Claudio
C
D000818:Animals; D017405:Herpes Simplex Virus Protein Vmw65 / Q000235:genetics / Q000378:metabolism; D006801:Humans; D051379:Mice; D008822:Mice, Transgenic; D046228:Microarray Analysis; D009119:Muscle Contraction / Q000502:physiology; D018482:Muscle, Skeletal / Q000166:cytology / Q000378:metabolism; D020388:Muscular Dystrophy, Duchenne / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D014018:Tissue Distribution; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D015854:Up-Regulation; D049411:Utrophin / Q000235:genetics / Q000378:metabolism*; D016335:Zinc Fingers / Q000235:genetics
D017405:Herpes Simplex Virus Protein Vmw65; D011993:Recombinant Fusion Proteins; D014157:Transcription Factors; D049411:Utrophin
Gene Ther. 2000 Feb;7(3):201-4
10694796
Nat Med. 2006 Oct;12(10):1147-50
16980968
Biochim Biophys Acta. 2007 Feb;1772(2):243-62
17064882
Biochem Cell Biol. 2007 Jun;85(3):358-65
17612630
Int Rev Cytol. 2004;240:1-30
15548414
Mol Pharmacol. 2004 Dec;66(6):1361-71
15340042
Neuromuscul Disord. 2005 Jun;15(6):437-49
15907291
Proc Natl Acad Sci U S A. 2005 Aug 16;102(33):11716-21
16081541
J Biol Chem. 2005 Oct 21;280(42):35588-97
16107335
Am J Physiol Heart Circ Physiol. 2005 Dec;289(6):H2373-8
16024571
Gene. 2006 Jan 17;366(1):27-38
16298089
Gene Ther. 2000 Jun;7(12):1076-83
10871758
J Mol Biol. 2000 Nov 3;303(4):489-502
11054286
Nat Genet. 2001 Feb;27(2):195-200
11175789
Oncogene. 2001 May 28;20(24):3116-27
11420728
J Biol Chem. 2001 Aug 3;276(31):29466-78
11340073
Am J Physiol Cell Physiol. 2001 Oct;281(4):C1300-9
11546668
Nat Genet. 2001 Dec;29(4):365-71
11726920
Annu Rev Biochem. 2001;70:313-40
11395410
Physiol Rev. 2002 Apr;82(2):291-329
11917091
Hum Mol Genet. 2002 Dec 15;11(26):3333-44
12471059
Bioinformatics. 2003 Jan 22;19(2):185-93
12538238
Curr Top Med Chem. 2003;3(6):645-57
12570857
Nat Rev Drug Discov. 2003 May;2(5):361-8
12750739
Nat Rev Drug Discov. 2003 May;2(5):379-90
12750741
Neuromuscul Disord. 2004 Jan;14(1):19-23
14659408
FEBS Lett. 2004 Feb 27;560(1-3):75-80
14988001
Biochem Cell Biol. 2004 Aug;82(4):428-36
15284895
Am J Physiol Lung Cell Mol Physiol. 2004 Sep;287(3):L569-76
15155269
Trends Mol Med. 2004 Oct;10(10):516-20
15464452
EMBO Rep. 2004 Sep;5(9):872-6
15470384
Neuron. 1991 Sep;7(3):499-508
1654951
Proc Natl Acad Sci U S A. 1993 Apr 15;90(8):3710-4
8475120
Proc Natl Acad Sci U S A. 1994 Nov 8;91(23):11163-7
7972027
Proc Natl Acad Sci U S A. 1994 Nov 8;91(23):11168-72
7972028
Nucleic Acids Res. 1996 May 1;24(9):1646-52
8649981
Nature. 1996 Nov 28;384(6607):308-9
8934508
Nature. 1996 Nov 28;384(6607):349-53
8934518
Curr Opin Struct Biol. 1997 Feb;7(1):117-25
9032060
FEBS Lett. 1997 Nov 3;417(1):71-4
9395077
Nat Genet. 1998 May;19(1):79-82
9590295
Nat Med. 1998 Dec;4(12):1441-4
9846586
Proc Natl Acad Sci U S A. 1999 Mar 16;96(6):2758-63
10077584
J Neuropathol Exp Neurol. 1999 Mar;58(3):235-44
10197815
Trends Mol Med. 2006 Mar;12(3):122-9
16443393
Assay Drug Dev Technol. 2006 Jun;4(3):317-31
16834537
Genome Res. 2006 Aug;16(8):1005-16
16809667
Nat Rev Mol Cell Biol. 2006 Oct;7(10):762-73
16971897
Nature. 2006 Nov 30;444(7119):574-9
17108972