scholarly journals The chicken skeletal muscle alpha-actin promoter is tissue specific in transgenic mice.

1989 ◽  
Vol 9 (9) ◽  
pp. 3785-3792 ◽  
Author(s):  
C J Petropoulos ◽  
M P Rosenberg ◽  
N A Jenkins ◽  
N G Copeland ◽  
S H Hughes
Keyword(s):  
Skeletal Muscle ◽  
Transgenic Mice ◽  
Striated Muscle ◽  
Transcriptional Start Site ◽  
Actin Gene ◽  
Tissue Specific ◽  
Adult Mice ◽  
Transgenic Lines ◽  
Alpha Actin ◽  
Chicken Skeletal Muscle

We have generated transgenic mouse lines that carry the promoter region of the chicken skeletal muscle alpha (alpha sk) actin gene linked to the bacterial chloramphenicol acetyltransferase (CAT) gene. In adult mice, the pattern of transgene expression resembled that of the endogenous alpha sk actin gene. In most of the transgenic lines, high levels of CAT activity were detected in striated muscle (skeletal and cardiac) but not in the other tissues tested. In striated muscle, transcription of the transgene was initiated at the normal transcriptional start site of the chicken alpha sk actin gene. The region from nucleotides -191 to +27 of the chicken alpha sk actin gene was sufficient to direct the expression of CAT in striated muscle of transgenic mice. These observations suggest that the mechanism of tissue-specific actin gene expression is well conserved in higher vertebrate species.

The chicken skeletal muscle alpha-actin promoter is tissue specific in transgenic mice

1989 ◽  
Vol 9 (9) ◽  
pp. 3785-3792
Author(s):  
C J Petropoulos ◽  
M P Rosenberg ◽  
N A Jenkins ◽  
N G Copeland ◽  
S H Hughes
Keyword(s):  
Skeletal Muscle ◽  
Transgenic Mice ◽  
Striated Muscle ◽  
Transcriptional Start Site ◽  
Actin Gene ◽  
Tissue Specific ◽  
Adult Mice ◽  
Transgenic Lines ◽  
Alpha Actin ◽  
Chicken Skeletal Muscle

We have generated transgenic mouse lines that carry the promoter region of the chicken skeletal muscle alpha (alpha sk) actin gene linked to the bacterial chloramphenicol acetyltransferase (CAT) gene. In adult mice, the pattern of transgene expression resembled that of the endogenous alpha sk actin gene. In most of the transgenic lines, high levels of CAT activity were detected in striated muscle (skeletal and cardiac) but not in the other tissues tested. In striated muscle, transcription of the transgene was initiated at the normal transcriptional start site of the chicken alpha sk actin gene. The region from nucleotides -191 to +27 of the chicken alpha sk actin gene was sufficient to direct the expression of CAT in striated muscle of transgenic mice. These observations suggest that the mechanism of tissue-specific actin gene expression is well conserved in higher vertebrate species.

scholarly journals CORP: Using transgenic mice to study skeletal muscle physiology

2020 ◽  
Vol 128 (5) ◽  
pp. 1227-1239
Author(s):  
C. Brooks Mobley ◽  
Ivan J. Vechetti ◽  
Taylor R. Valentino ◽  
John J. McCarthy
Keyword(s):  
Skeletal Muscle ◽  
Transgenic Mice ◽  
Research Program ◽  
Gene Function ◽  
Cell Biology ◽  
Muscle Physiology ◽  
Basic Information ◽  
Adult Skeletal Muscle ◽  
Tissue Specific ◽  
Muscle Research

The development of tissue-specific inducible transgenic mice has provided a powerful tool to study gene function and cell biology in almost any tissue of interest at any given time within the animal’s life. The purpose of this review is to describe how to use two different inducible transgenic systems, the Cre-loxP system and the Tet-ON/OFF system, that can be used to study skeletal muscle physiology. Myofiber- and satellite cell-specific Cre-loxP transgenic mice are described as is how these mice can be used to knockout a gene of interest or to deplete satellite cells in adult skeletal muscle, respectively. A myofiber-specific Tet-ON system is described as is how such mice can be used to overexpress a gene of interest or to label myonuclei. How to effectively breed and genotype the transgenic mice are also described in detail. The hope is this review will provide the basic information necessary to facilitate the incorporation of tissue-specific inducible transgenic mice into a skeletal muscle research program.

scholarly journals Tissue-specific and developmentally regulated expression of a chimeric actin-globin gene in transgenic mice.

1986 ◽  
Vol 6 (7) ◽  
pp. 2624-2631 ◽  
Author(s):  
M Shani
Keyword(s):  
Transgenic Mice ◽  
Dna Sequences ◽  
Globin Gene ◽  
Chimeric Gene ◽  
Actin Gene ◽  
Muscle Actin ◽  
Developmentally Regulated ◽  
Tissue Specific ◽  
Regulated Expression ◽  
Muscle Actin Gene

A chimeric plasmid containing about 2/3 of the rat skeletal muscle actin gene plus 730 base pairs of its 5' flanking sequences fused to the 3' end of a human embryonic globin gene (D. Melloul, B. Aloni, J. Calvo, D. Yaffe, and U. Nudel, EMBO J. 3:983-990, 1984) was inserted into mice by microinjection into fertilized eggs. Eleven transgenic mice carrying the chimeric gene with or without plasmid pBR322 DNA sequences were identified. The majority of these mice transmitted the injected DNA to about 50% of their progeny. However, in transgenic mouse CV1, transmission to progeny was associated with amplification or deletion of the injected DNA sequences, while in transgenic mouse CV4 transmission was distorted, probably as a result of insertional mutagenesis. Tissue-specific expression was dependent on the removal of the vector DNA sequences from the chimeric gene sequences prior to microinjection. None of the transgenic mice carrying the chimeric gene together with plasmid pBR322 sequences expressed the introduced gene in striated muscles. In contrast, the six transgenic mice carrying the chimeric gene sequences alone expressed the inserted gene specifically in skeletal and cardiac muscles. Moreover, expression of the chimeric gene was not only tissue specific, but also developmentally regulated. Similar to the endogenous skeletal muscle actin gene, the chimeric gene was expressed at a relatively high level in cardiac muscle of neonatal mice and at a significantly lower level in adult cardiac muscle. These results indicate that the injected DNA included sufficient cis-acting control elements for its tissue-specific and developmentally regulated expression in transgenic mice.

Juxtaposition of the HPFH2 Enhancer Is Not Sufficient To Reactivate the g-Globin Gene in Adult Erythropoiesis in bYAC Transgenic Mice.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 1215-1215
Author(s):  
Ping Xiang ◽  
Xin Ye ◽  
Hemei Han ◽  
Mary Stafford ◽  
Stamatoyannopoulos George ◽  
...  
Keyword(s):  
Transgenic Mice ◽  
Developmental Stages ◽  
Globin Gene ◽  
Fetal Hemoglobin ◽  
Upstream Region ◽  
Rnase Protection ◽  
Adult Mice ◽  
Gene Reactivation ◽  
Transgenic Lines ◽  
High Level

Abstract Hereditary persistence of fetal hemoglobin (HPFH) is characterized by high-level expression of the g globin gene in adult patients. HPFH2 contains a genetic deletion of approximately 83.5 kb from the middle of intron 2 of the yb gene to the region 66 kb 3′ to the b gene. The deletion juxtaposes an enhancer located downstream to the 3′ breakpoint to the vicinity of the g gene. To understand the reactivation mechanism of this fetal stage specific globin, we introduced the deletion into a 213 kb b-YAC. Four transgenic lines bearing 1–3 intact copies of the YAC construct were established. Globin gene expression at different developmental stages was measured by RNase protection assays. In the HPFH2 transgenic mice the e and g genes were expressed at high levels similar to the wild type YAC mice in embryonic and fetal stages. Unexpectedly, the g gene was completely silenced in the adult mice. The failure of g gene reactivation by juxtaposition of the HPFH2 enhancer contradicts the results reported by the Forget lab (Mol. Cell. Biol.17:2076–2089, 1997) and the Strouboulis lab (Blood102:3412–3419, 2003). The discrepancy could be due to the differences between the constructs used in the different labs. The construct used in the Forget lab was a cosmid containing the mLCR linked a 13 kb Gg and Ag fragment. The cosmid used by the Strouboulis lab contained the 22 kb LCR and a 5 kb Ag fragment. The bYAC construct we used in this study contains the whole b globin locus and spanned from about 40 kb 5′ to the e gene to 100 kb 3′ to the b gene. The failure of g gene reactivation in the HPFH2 YAC mice suggests that additional elements, which are missing in the 213 kb bYAC construct, are involved in reactivation of the g gene in the HPFH patients. Based on data from literature (Bender et al., Mol. Cell5:387–393, 2000; Forrester et al., Genes & Dev.4:1637–1649, 1990) and our own studies (Ping et al., this meeting) we speculate that the 200 kb upstream region is the candidate for harboring this activity.

scholarly journals GGeneration of an efficient artificial promoter of bovine skeletal muscle alpha-actin gene (ACTA1) through addition of cis-acting element

2015 ◽  
Vol 20 (1) ◽  
Author(s):  
Qian Hu ◽  
Huili Tong ◽  
Dandan Zhao ◽  
Yunkao Cao ◽  
Weiwei Zhang ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Core Promoter ◽  
Genetically Engineered ◽  
Actin Gene ◽  
Base Pairs ◽  
Cis Acting ◽  
The Core ◽  
Binding Factor ◽  
Bovine Skeletal Muscle ◽  
Alpha Actin

AbstractThe promoter of skeletal muscle α-actin gene (ACTA1) is highly muscle specific. The core of the bovine ACTA1 promoter extends from +29 to −233, about 262 base pairs (bp), which is sufficient to activate transcription in bovine muscle satellite cells. In this study, analysis by PCR site-specific mutagenesis showed that the cis-acting element SRE (serum response element binding factor) was processed as a transcriptional activator. In order to enhance the bovine ACTA1 promoter’s activity, we used a strategy to modify it. We cloned a fragment containing three SREs from the promoter of ACTA1, and then one or two clones were linked upstream of the core promoter (262 bp) of ACTA1. One and two clones increased the activity of the ACTA1 promoter 3-fold and 10-fold, respectively, and maintained muscle tissue specificity. The modified promoter with two clones could increase the level of ACTA1 mRNA and protein 4-fold and 1.1-fold, respectively. Immunofluorescence results showed that green fluorescence of ACTA1 increased. Additionally, the number of total muscle microfilaments increased. These genetically engineered promoters might be useful for regulating gene expression in muscle cells and improving muscle mass in livestock.

Differential expression of the myocyte enhancer factor 2 family of transcription factors in development: the cardiac factor BBF-1 is an early marker for cardiogenesis

1994 ◽  
Vol 14 (8) ◽  
pp. 5130-5138
Author(s):  
S Goswami ◽  
P Qasba ◽  
S Ghatpande ◽  
S Carleton ◽  
A K Deshpande ◽  
...  
Keyword(s):  
Dna Binding ◽  
Cardiac Muscle ◽  
Differential Expression ◽  
Actin Gene ◽  
Myocyte Enhancer Factor 2 ◽  
Tissue Specific ◽  
Myocyte Enhancer Factor ◽  
Gene Transcripts ◽  
Alpha Actin ◽  
Enhancer Factor

In the present study, we have used single chicken blastoderms of defined early developmental stages, beginning with the prestreak stage, stage 1 (V. Hamburger and H. L. Hamilton, J. Morphol. 88:49-92, 1951), to analyze the onset of cardiac myogenesis by monitoring the appearance of selected cardiac muscle tissue-specific gene transcripts and the functional expression of the myocyte enhancer factor 2 (MEF-2) proteins. Using gene-specific oligonucleotide primers in reverse transcriptase PCR assay, we have demonstrated that the cardiac myosin light-chain 2 (MLC2) and alpha-actin gene transcripts appear as early as stage 5, i.e., immediately after the cardiogenic fate assignment at stage 4. Consistent with this observation is the developmental expression pattern of DNA-binding activity of BBF-1, a cardiac muscle-specific member of the MEF-2 protein family, which also begins at stage 5 prior to MEF-2. Differential expression of DNA-binding complexes is also observed with another AT-rich DNA sequence (CArG box) as probe, but the binding pattern with the ubiquitous TATA-binding proteins remains unchanged during the same developmental period. Thus, the cardiogenic commitment and differentiation of the precardiac mesoderm, as exemplified by the appearance of cardiac MEF-2, MLC2, and alpha-actin gene products, occur earlier than previously thought and appear to be closely linked. The onset of skeletal myogenic program follows that of the cardiogenic program with the appearance of skeletal MLC2 at stage 8. We also observed that mRNA for the MEF-2 family of proteins appears as early as stage 2 and that for CMD-1, the chicken counterpart of MyoD, appears at stage 5. The temporal separation of activation of cardiac and skeletal MLC2 genes, which appears immediately after the respective fate assignments, and those of cardiac MEF-2 and CMD-1, which occur before, are consistent with the established appearance of the myogenic programs and with the acquisition pattern of the two tissue-specific morphological characteristics in the early embryo. The preferential appearance of BBF-1 activity in precardiac moesderm, relative to that of MEF-2, indicates that these two protein factors are distinct members of the MEF-2 family and provides a compelling argument in support of the potential role of BBF-1 as a regulator of the cardiogenic cell lineage determination, while cardiac MEF-2 might be involved in maintenance of the cardiac differentiative state.

Functionally distinct elements are required for expression of the AMPD1 gene in myocytes

1993 ◽  
Vol 13 (9) ◽  
pp. 5854-5860
Author(s):  
T Morisaki ◽  
E W Holmes
Keyword(s):  
Skeletal Muscle ◽  
Fiber Type ◽  
Transcriptional Start Site ◽  
Protein S ◽  
Muscle Fiber Types ◽  
Binding Motif ◽  
Fiber Types ◽  
Specific Expression ◽  
Tissue Specific ◽  
Ampd1 Gene

AMP deaminase (AMPD) is an enzyme found in all eukaryotic cells. Tissue-specific and stage-specific isoforms of this enzyme are found in vertebrates, and expression of these different isoforms is determined by selective expression of the multiple genes. The AMPD1 gene is expressed predominantly in skeletal muscle, in which transcript abundance is controlled by stage-specific and fiber type-specific signals. This enzyme activity is presumed to be important in skeletal muscle because a metabolic myopathy develops in individuals with an inherited deficiency of AMPD1. In the present study, cis- and trans-acting factors that control expression of AMPD1 have been identified. Two cis-acting elements located within 100 nucleotides of the transcriptional start site are required for muscle-specific expression of AMPD1. One element (-100 to -79) behaves like a tissue-specific enhancer, and it interacts with protein(s) found predominantly in nuclei of myoblasts and myotubes. This element is similar in sequence to an MEF2 binding motif, and it contains an A/T core that is essential for enhancer activity and binding of a nuclear protein(s). The second element (-60 to -40) has properties of a stage-specific promoter in that it is essential for muscle-specific expression of the AMPD1 promoter, does not confer muscle-specific expression on a heterologous promoter construct, and interacts with a protein(s) restricted to nuclei of differentiated myotubes. Interaction between these functionally distinct elements may be required for regulating the expression of AMPD1 during myocyte differentiation and in different muscle fiber types.

Muscle disease caused by mutations in the skeletal muscle alpha-actin gene (ACTA1)

2003 ◽  
Vol 13 (7-8) ◽  
pp. 519-531 ◽  
Author(s):  
John C. Sparrow ◽  
Kristen J. Nowak ◽  
Hayley J. Durling ◽  
Alan H. Beggs ◽  
Carina Wallgren-Pettersson ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Muscle Disease ◽  
Actin Gene ◽  
Alpha Actin

Severe nemaline myopathy caused by mutations of the stop codon of the skeletal muscle alpha actin gene (ACTA1)

2006 ◽  
Vol 16 (9-10) ◽  
pp. 541-547 ◽  
Author(s):  
William Wallefeld ◽  
Sabine Krause ◽  
Kristen J. Nowak ◽  
Danielle Dye ◽  
Rita Horváth ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Stop Codon ◽  
Nemaline Myopathy ◽  
Actin Gene ◽  
Alpha Actin
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