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                   <span class='card-content'>1 Abstract</span>
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                   <span class='card-content'>1 Experiments in iGEM</span>
 
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                   <span class='card-content'>2 Introduction</span>
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                   <span class='card-content'>2 Experiments before iGEM</span>
 
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                   <span class='card-content'>3 Results</span>
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<h2 class="inner-h">1 Experiments in iGEM</h2>
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Chronic PKA phosphorylation of RyR2 has been shown to increased diastolic SR Ca<sup>2+</sup> leak and lead to cardiac dysfunction. Since the change of phosphorylation level of RyR2 is a biomarker of failing heart, we attempted to verify the hypothesis that intracellular gene delivery of a RyR2 targeting phosphorylation site-specific nanobody could preserve contractility of failing myocardium. In present study, we acquired the RyR2-specific nanobodies from a phage display library which are variable domains of camellidae heavy chain-only antibodies (VHH). One of the monoclonal nanobodies, AR185, inhibiting RyR2 phosphorylation in an in vitro assay was then chosen for further investigation. We investigated the potential of adeno-associated virus (AAV)-9-mediated cardiac expression of AR185 against post-ischemic heart failure. Adeno-associated virus gene delivery elevated the intracellular expression AR185 protein in the ischemic heart failure model of rats, and this treatment normalized the systolic and diastolic dysfunction of the failing myocardium in vivo and in vitro by reversing myocardial Ca<sup>2+</sup> handling. Furthermore, AR185 gene transfer to failing cardiomyocytes reduced the frequency of sarcoplasmic reticulum (SR) calcium leak, thereby restoring the attenuated intracellular calcium transients and SR calcium load. Moreover, AR185 gene transfer inhibited PKA phosphorylation of RyR2 in failing cardiomyocytes. Our results provided strong pre-clinical experimental evidence of the cardiac expression of RyR2 nanobody with AAV9 vectors as a promising therapeutic strategy for ischemic heart failure.
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<h2>Wanna contact us?</h2>
 
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Latest revision as of 15:58, 20 November 2018

1 Experiments in iGEM
2 Experiments before iGEM
3 Basic experiments and protocol

Experiments

1 Experiments in iGEM

2 Experiments before iGEM

3 Basic experiments and protocol

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