Utrophin Up-Regulation by an Artificial Transcription Factor in Transgenic Mice
Publication Date
August 22, 2007
Journal
PLOS ONE
Authors
Elisabetta Mattei, Nicoletta Corbi, Maria Grazia Di Certo, Georgios Strimpakos, et al
Volume
2
Issue
8
Pages
e774
DOI
http://doi.org/10.1371/journal.pone.0000774
Publisher URL
http://journals.plos.org/plosone/article?id=10.1371%2Fjournal.pone.0000774
PubMed
http://www.ncbi.nlm.nih.gov/pubmed/17712422
PubMed Central
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1942121
Europe PMC
http://europepmc.org/abstract/MED/17712422
Web of Science
000207455300022
Scopus
40749102811
Mendeley
http://www.mendeley.com/research/utrophin-upregulation-artificial-transciption-factor-transgenic-mice
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Mendeley | Further Information

{"title"=>"Utrophin up-regulation by an artificial transciption factor in transgenic mice", "type"=>"journal", "authors"=>[{"first_name"=>"Elisabetta", "last_name"=>"Mattei", "scopus_author_id"=>"7004257300"}, {"first_name"=>"Nicoletta", "last_name"=>"Corbi", "scopus_author_id"=>"6603251655"}, {"first_name"=>"Maria Grazia", "last_name"=>"Di Certo", "scopus_author_id"=>"6507925372"}, {"first_name"=>"Georgios", "last_name"=>"Strimpakos", "scopus_author_id"=>"23977078800"}, {"first_name"=>"Cinzia", "last_name"=>"Severini", "scopus_author_id"=>"7005669520"}, {"first_name"=>"Annalisa", "last_name"=>"Onori", "scopus_author_id"=>"8760586100"}, {"first_name"=>"Agata", "last_name"=>"Desantis", "scopus_author_id"=>"16645148400"}, {"first_name"=>"Valentina", "last_name"=>"Libri", "scopus_author_id"=>"8069638900"}, {"first_name"=>"Serena", "last_name"=>"Buontempo", "scopus_author_id"=>"23995181900"}, {"first_name"=>"Aristide", "last_name"=>"Floridi", "scopus_author_id"=>"7103094412"}, {"first_name"=>"Maurizio", "last_name"=>"Fanciulli", "scopus_author_id"=>"7006373109"}, {"first_name"=>"Dilair", "last_name"=>"Baban", "scopus_author_id"=>"6602934893"}, {"first_name"=>"Kay E.", "last_name"=>"Davies", "scopus_author_id"=>"35414895000"}, {"first_name"=>"Claudio", "last_name"=>"Passananti", "scopus_author_id"=>"6701533321"}], "year"=>2007, "source"=>"PLoS ONE", "identifiers"=>{"isbn"=>"1932-6203 (Electronic) 1932-6203 (Linking)", "doi"=>"10.1371/journal.pone.0000774", "pui"=>"351385870", "sgr"=>"40749102811", "scopus"=>"2-s2.0-40749102811", "pmid"=>"17712422", "issn"=>"19326203"}, "id"=>"a7b14ff6-4c87-3a4b-945b-cc10c022cf27", "abstract"=>"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.", "link"=>"http://www.mendeley.com/research/utrophin-upregulation-artificial-transciption-factor-transgenic-mice", "reader_count"=>21, "reader_count_by_academic_status"=>{"Professor > Associate Professor"=>2, "Researcher"=>8, "Student > Doctoral Student"=>1, "Student > Ph. D. Student"=>4, "Other"=>2, "Student > Master"=>1, "Professor"=>3}, "reader_count_by_user_role"=>{"Professor > Associate Professor"=>2, "Researcher"=>8, "Student > Doctoral Student"=>1, "Student > Ph. D. Student"=>4, "Other"=>2, "Student > Master"=>1, "Professor"=>3}, "reader_count_by_subject_area"=>{"Biochemistry, Genetics and Molecular Biology"=>3, "Agricultural and Biological Sciences"=>13, "Medicine and Dentistry"=>3, "Chemistry"=>1, "Immunology and Microbiology"=>1}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>3}, "Chemistry"=>{"Chemistry"=>1}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>13}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>3}}, "reader_count_by_country"=>{"Egypt"=>1, "United Kingdom"=>2, "Italy"=>1, "Slovenia"=>1, "Chile"=>1}, "group_count"=>1}

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Scopus | Further Information

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  • {"files"=>["https://ndownloader.figshare.com/files/946159"], "description"=>"<p>A: Schematic representation of the Vp16-Jazz gene in the pMex-vector, used to generate transgenic mice. The 9 base pair long Vp16-Jazz DNA target sequence is indicated. B: The nucleotide sequence of the mouse utrophin promoter A. The Vp16-Jazz DNA target sequence is indicated in bold characters and underlined. The main transcription factor binding sites present in this promoter region are indicated.</p>", "links"=>[], "tags"=>["dna"], "article_id"=>616498, "categories"=>["Molecular Biology", "Biotechnology", "Neuroscience"], "users"=>["Elisabetta Mattei", "Nicoletta Corbi", "Maria Grazia Di Certo", "Georgios Strimpakos", "Cinzia Severini", "Annalisa Onori", "Agata Desantis", "Valentina Libri", "Serena Buontempo", "Aristide Floridi", "Maurizio Fanciulli", "Dilair Baban", "Kay E. Davies", "Claudio Passananti"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0000774.g001", "stats"=>{"downloads"=>3, "page_views"=>13, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Vp16_Jazz_and_its_DNA_target_sequence_/616498", "title"=>"Vp16-Jazz and its DNA target sequence.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 12:02:47"}
  • {"files"=>["https://ndownloader.figshare.com/files/946371"], "description"=>"<p>A: Western Blot analysis of total proteins extracted from the skeletal muscle of wild type (wt) and transgenic mice derived from two different founders (tg9 and tg41). The expression of Vp16-Jazz transgene was monitored by the anti-myc monoclonal 9E10 antibody. Detection of α−tubulin was used to normalize the amount of proteins. B: Western Blot analysis of total proteins extracted from the skeletal muscle, heart and brain of transgenic mice from families tg9 and tg41.</p>", "links"=>[], "tags"=>["vp16-jazz", "transgenic"], "article_id"=>616707, "categories"=>["Molecular Biology", "Biotechnology", "Neuroscience"], "users"=>["Elisabetta Mattei", "Nicoletta Corbi", "Maria Grazia Di Certo", "Georgios Strimpakos", "Cinzia Severini", "Annalisa Onori", "Agata Desantis", "Valentina Libri", "Serena Buontempo", "Aristide Floridi", "Maurizio Fanciulli", "Dilair Baban", "Kay E. Davies", "Claudio Passananti"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0000774.g002", "stats"=>{"downloads"=>0, "page_views"=>22, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Muscle_specific_expression_of_Vp16_Jazz_in_transgenic_mice_/616707", "title"=>"Muscle specific expression of Vp16-Jazz in transgenic mice.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 12:03:57"}
  • {"files"=>["https://ndownloader.figshare.com/files/946639"], "description"=>"<p>Immunohistochemistry of Tibialis Anterior (TA) muscle derived from wt (panel A) and transgenic mice tg9 (panel B) stained with anti-utrophin antibody. Nuclei are counterstained with Hoechst 33258. C: TA from wt and transgenic mice tg9 were co-stained with anti-utrophin antibody and the α-bungarotoxin-Alexa Fluor to visualize the acetylcholine receptor (AChR) at the neuromuscular junctions. The anti-utrophin monoclonal antibody reveals an extra-synaptic distribution of utrophin only in transgenic mice. D: Relationship between contractile response (g) and stimulation frequency (Volts) in diaphragm and EDL muscle preparations obtained from wt, tg9 and tg41 mice. E: Scatter plots of the DEG with natural log transformed expression values averaged over the 4 replicates. The x–axis reports the mean of the replicates of transgenic mice, while the y-axis is the mean across the replicates of the control wt mice. Expression values are colour coded with red representing up-regulated genes in tg9/tg41 and green down-regulated genes compared to wild-type and lines on the graph set at 2-folds differential expression.</p>", "links"=>[], "tags"=>["vp16-jazz", "transgenic"], "article_id"=>616981, "categories"=>["Molecular Biology", "Biotechnology", "Neuroscience"], "users"=>["Elisabetta Mattei", "Nicoletta Corbi", "Maria Grazia Di Certo", "Georgios Strimpakos", "Cinzia Severini", "Annalisa Onori", "Agata Desantis", "Valentina Libri", "Serena Buontempo", "Aristide Floridi", "Maurizio Fanciulli", "Dilair Baban", "Kay E. Davies", "Claudio Passananti"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0000774.g004", "stats"=>{"downloads"=>2, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effects_of_Vp16_Jazz_in_transgenic_mice_/616981", "title"=>"Effects of Vp16-Jazz in transgenic mice.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 12:06:01"}
  • {"files"=>["https://ndownloader.figshare.com/files/946516"], "description"=>"<p>A: Vp16-Jazz chromatin immunoprecipitation, performed in skeletal muscle derived from wt mice and transgenic mice (family tg9) using myc monoclonal antibody/protein G-agarose beads or protein G-agarose beads as a control (no-Ab). Immunoprecipitates from each sample were analyzed for the presence of utrophin promoter by PCR. A sample representing linear amplification of the total input chromatin (input) was included (lane 1). As control, samples from transgenic mice were also tested for the presence of dystrophin promoter sequence. B: Real-time PCR analysis of the utrophin gene expression rate in Vp16-Jazz transgenic mice (tg9 and tg41) and control wt mice. The gene expression ratio between utrophin and β-glucoronidase (GUS) and β2-microglobulin (β2M) is shown as means±S.D. from three independent experiments performed in triplicate. C: Western blot of total protein extracts derived from skeletal muscle and heart from wt mice and Vp16-Jazz transgenic mice (tg9 and tg41) incubated with monoclonal antibody against utrophin. The same membrane was incubated with anti-α-tubulin monoclonal antibody for loading normalization. D: Relative utrophin expression of wt and transgenic mice (tg9 and tg41) was determined by densitometric analysis. E: Total protein extracts from skeletal muscle of wt and transgenic mice (tg9 and tg41) were subjected to immunoblotting to detect the expression levels of the dystrophin and α-sarcoglycan proteins. The anti-α-tubulin and anti-myc antibodies were used to normalize the protein content and to test the Vp16-Jazz transgene expression respectively.</p>", "links"=>[], "tags"=>["utrophin"], "article_id"=>616854, "categories"=>["Molecular Biology", "Biotechnology", "Neuroscience"], "users"=>["Elisabetta Mattei", "Nicoletta Corbi", "Maria Grazia Di Certo", "Georgios Strimpakos", "Cinzia Severini", "Annalisa Onori", "Agata Desantis", "Valentina Libri", "Serena Buontempo", "Aristide Floridi", "Maurizio Fanciulli", "Dilair Baban", "Kay E. Davies", "Claudio Passananti"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0000774.g003", "stats"=>{"downloads"=>1, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Vp16_Jazz_and_utrophin_up_regulation_/616854", "title"=>"Vp16-Jazz and utrophin up-regulation.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 12:05:13"}

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