Strength and muscle specificity of a compact promoter derived from the slow troponin I gene in the context of episomal (gutless adenovirus) and integrating (lentiviral) vectors

2012 ◽  
Vol 14 (12) ◽  
pp. 746-760 ◽  
Author(s):  
Marc-André Robert ◽  
Yuanbang Lin ◽  
Mehdi Bendjelloul ◽  
Yue Zeng ◽  
Sofien Dessolin ◽  
...  
Keyword(s):  
Troponin I ◽  
Lentiviral Vectors ◽  
Muscle Specificity ◽  
I Gene

Functional Consequences of the Mutations in Human Cardiac Troponin I Gene Found in Familial Hypertrophic Cardiomyopathy

2001 ◽  
Vol 33 (12) ◽  
pp. 2095-2107 ◽  
Author(s):  
Fumi Takahashi-Yanaga ◽  
Sachio Morimoto ◽  
Keita Harada ◽  
Reiko Minakami ◽  
Fumie Shiraishi ◽  
...  
Keyword(s):  
Hypertrophic Cardiomyopathy ◽  
Cardiac Troponin ◽  
Troponin I ◽  
Cardiac Troponin I ◽  
Familial Hypertrophic Cardiomyopathy ◽  
Functional Consequences ◽  
I Gene

scholarly journals Troponin I gene expression during human cardiac development and in end-stage heart failure.

1993 ◽  
Vol 72 (5) ◽  
pp. 932-938 ◽  
Author(s):  
S Sasse ◽  
N J Brand ◽  
P Kyprianou ◽  
G K Dhoot ◽  
R Wade ◽  
...  
Keyword(s):  
Gene Expression ◽  
Heart Failure ◽  
Troponin I ◽  
Cardiac Development ◽  
End Stage Heart Failure ◽  
End Stage ◽  
I Gene

scholarly journals In vivo recognition of a vertebrate mini-exon as an exon-intron-exon unit.

1993 ◽  
Vol 13 (5) ◽  
pp. 2677-2687 ◽  
Author(s):  
D A Sterner ◽  
S M Berget
Keyword(s):  
Splice Site ◽  
Rna Splicing ◽  
Troponin I ◽  
Exon Skipping ◽  
Splice Sites ◽  
Small Vertebrate ◽  
Upstream Exon ◽  
I Gene

Very small vertebrate exons are problematic for RNA splicing because of the proximity of their 3' and 5' splice sites. In this study, we investigated the recognition of a constitutive 7-nucleotide mini-exon from the troponin I gene that resides quite close to the adjacent upstream exon. The mini-exon failed to be included in spliced RNA when placed in a heterologous gene unless accompanied by the upstream exon. The requirement for the upstream exon disappeared when the mini-exon was internally expanded, suggesting that the splice sites bordering the mini-exon are compatible with those of other constitutive vertebrate exons and that the small size of the exon impaired inclusion. Mutation of the 5' splice site of the natural upstream exon did not result in either exon skipping or activation of a cryptic 5' splice site, the normal vertebrate phenotypes for such mutants. Instead, a spliced RNA accumulated that still contained the upstream intron. In vitro, the mini-exon failed to assemble into spliceosome complexes unless either internally expanded or accompanied by the upstream exon. Thus, impaired usage of the mini-exon in vivo was accompanied by impaired recognition in vitro, and recognition of the mini-exon was facilitated by the presence of the upstream exon in vivo and in vitro. Cumulatively, the atypical in vivo and in vitro properties of the troponin exons suggest a mechanism for the recognition of this mini-exon in which initial recognition of an exon-intron-exon unit is followed by subsequent recognition of the intron.

scholarly journals Heterogeneity of Clinical Manifestation of Hypertrophic Cardiomyopathy Caused by Deletion of Lysine 183 in Cardiac Troponin I Gene

2010 ◽  
Vol 51 (3) ◽  
pp. 214-217 ◽  
Author(s):  
Akira Funada ◽  
Eiichi Masuta ◽  
Noboru Fujino ◽  
Kenshi Hayashi ◽  
Hidekazu Ino ◽  
...  
Keyword(s):  
Hypertrophic Cardiomyopathy ◽  
Clinical Manifestation ◽  
Cardiac Troponin ◽  
Troponin I ◽  
Cardiac Troponin I ◽  
I Gene

Abstract 1118: Novel Lentiviral Vectors Bearing the Cardiac-Specific Promoter of the Sodium-Calcium Exchanger (NCX1) Report Cardiogenic Differentiation in Stem Cells

Circulation ◽  
2007 ◽  
Vol 116 (suppl_16) ◽  
Author(s):  
Andreas S Barth ◽  
Eddy Kizana ◽  
John Terrovitis ◽  
Peihong Dong ◽  
Rachel R Smith ◽  
...  
Keyword(s):  
Stem Cells ◽  
Troponin I ◽  
Lentiviral Vectors ◽  
Neonatal Rat ◽  
Luciferase Reporter ◽  
Sodium Calcium Exchanger ◽  
Α Subunit ◽  
Sodium Calcium ◽  
Cardiogenic Differentiation

Background: Cardiosphere-derived resident cardiac stem cells (CDCs) are readily isolated from adult hearts and confer functional benefit in animal models of heart failure. To study cardiogenic differentiation in CDCs, we developed a method to genetically label and selectively enrich for cells that have differentiated down the cardiomyocyte lineage. Methods and Results: Lentiviral vectors achieved significantly higher transduction efficiency (>90%) and longer-term transgene expression in human CDCs than any of the nine adeno-associated viral serotypes tested (AAV 1– 6, 2.5, 8, 9). To define the best reporter of cardiogenic differentiation, five cardiac-specific promoters (sodium-calcium exchanger [NCX1], L-type calcium channel α-subunit, α-myosin heavy chain, troponin I and myosin ventricular light chain) were subcloned upstream of GFP in lentiviral vectors. Cardiac specificity was assessed by transducing neonatal rat ventricular myocytes (NRVMs), HEK293, cardiac fibroblasts, skeletal myoblasts, endothelial and vascular smooth muscle cells with each vector and then measuring GFP fluorescence by flow cytometry. The cardiac NCX1-promoter conveyed the highest degree of cardiac specificity, with expression limited to NRVMs. At a multiplicity of infection of 35, NCX1-GFP vectors did not affect cell death or apoptosis of CDCs. Expression persisted for up to 6 months in vitro . NCX1-GFP positive CDCs subpopulations, were FACS-sorted and expanded in vitro , demonstrating enhanced expression of a variety of cardiac markers by real-time PCR. The utility of lentiviral vectors bearing the cardiac NCX1-promoter was further exemplified by treating CDCs with cardiogenic sulfonylhydrazones (SHZ). SHZ potently activated a NCX1-firefly luciferase reporter (211 ± 21% greater than control cells) and upregulated cardiac transcripts (including BNP, troponin I, α-tropomyosin, GATA-4). Conclusion: Lentiviral vectors carrying the NCX1 promoter represent a useful tool for genetically marking stem cells and their progeny that have differentiated down the cardiomyocyte lineage. The ability to selectively enrich for cardiomyocytes and their precursors has potential relevance for the development of cell-based therapies.

Complex regulation of the muscle-specific contractile protein (troponin I) gene

1987 ◽  
Vol 7 (9) ◽  
pp. 3065-3075
Author(s):  
S F Konieczny ◽  
C P Emerson
Keyword(s):  
Transcriptional Activation ◽  
Troponin I ◽  
Protein Gene ◽  
Regulatory Elements ◽  
Developmental Expression ◽  
Contractile Protein ◽  
Upstream Region ◽  
Myoblast Differentiation ◽  
Quantitative Expression ◽  
I Gene

A cloned quail troponin I contractile protein gene, stably transfected into a mouse myogenic cell line, exhibits appropriate developmental activation and quantitative expression during myoblast differentiation. Deletion mutagenesis analyses reveal that the troponin I gene has two distinct cis regulatory elements required for its developmental expression, as measured by mRNA accumulation and nuclear runoff transcription assays. One element in the 5' flanking region is required for maximum quantitative expression, and a second larger regulatory element (1.5 kilobases) within the first intron is responsible for differentiation-specific transcription. The upstream region is highly sensitive to negative repression by interaction with pBR322 sequences. The larger intragenic region retains some activity when moved to the 5' and 3' flanking regions and when inverted but is maximally active in its native intragenic site. The concerted activities of these two regulatory regions produce a 100- to 200-fold transcriptional activation during myoblast differentiation. The conserved 5' exon-intron organization of troponin I and other contractile protein genes suggests a possible mechanism by which intragenic control elements coordinate contractile protein gene regulation during skeletal myogenesis.

scholarly journals Structure, evolution, and regulation of a fast skeletal muscle troponin I gene.

1985 ◽  
Vol 82 (23) ◽  
pp. 8080-8084 ◽  
Author(s):  
A. S. Baldwin ◽  
E. L. Kittler ◽  
C. P. Emerson
Keyword(s):  
Skeletal Muscle ◽  
Troponin I ◽  
Structure Evolution ◽  
Fast Skeletal Muscle ◽  
I Gene

scholarly journals Regulation of the rat cardiac troponin I gene by the transcription factor GATA-4

1997 ◽  
Vol 322 (2) ◽  
pp. 393-401 ◽  
Author(s):  
Anne M. MURPHY ◽  
W. Reid THOMPSON ◽  
Ling Fan PENG ◽  
Lawrence JONES
Keyword(s):  
Transcription Factor ◽  
Cardiac Troponin ◽  
Troponin I ◽  
Cardiac Cells ◽  
Cardiac Troponin I ◽  
Luciferase Expression ◽  
Specific Gene ◽  
Upstream Sequence ◽  
I Gene

Troponin I is a thin-filament contractile protein expressed in striated muscle. There are three known troponin I genes which are expressed in a muscle-fibre-type-specific manner in mature animals. Although the slow skeletal troponin I isoform is expressed in fetal and neonatal heart, the cardiac isoform is restricted in its expression to the myocardium at all developmental stages. To study the regulation of this cardiac-specific and developmentally regulated gene in vitro, the rat cardiac troponin I gene was cloned. Transient transfection assays were performed with troponin I–luciferase fusion plasmids to characterize the regulatory regions of the gene. Proximal regions of the upstream sequence were sufficient to support high levels of expression of the reporter gene in cardiocytes and relatively low levels in other cell types. The highest luciferase activity in the cardiocytes was noted with a plasmid that included the region spanning -896 to +45 of the troponin I genomic sequence. Co-transfection of GATA-4, a recently identified cardiac transcription factor, with troponin I–luciferase constructs permitted high levels of luciferase expression in non-cardiac cells. Electrophoretic mobility-shift assays demonstrated specific binding of GATA-4 to oligonucleotides representative of multiple sites of the troponin I sequence. Mutation of a proximal GATA-4 DNA-binding site decreased transcriptional activation in transfected cardiocytes. These results indicate that the proximal cardiac troponin I sequence is sufficient to support high levels of cardiac-specific gene expression and that the GATA-4 transcription factor regulates troponin I–luciferase expression in vitro.

scholarly journals Analysis of troponin I gene polymorphisms and meat quality in Mongcai pigs

2012 ◽  
Vol 42 (3) ◽  
Author(s):  
NT Ngu ◽  
NTH Nhan
Keyword(s):  
Meat Quality ◽  
Gene Polymorphisms ◽  
Troponin I ◽  
I Gene
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