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Updated: May 11, 2025
Preparation of rAAV9 to Overexpress or Knockdown Genes in Mouse Hearts
Published on: December 17, 2016
Protocol for assessing regulatory elements in murine heart using an AAV9-based massively parallel reporter assay
Kaiyu Wang1, Meng Meng2, Minghong Leng2
1Department of Pharmacology, Joint Laboratory of Guangdong-Hong Kong Universities for Vascular Homeostasis and Diseases, School of Medicine, Southern University of Science and Technology, Shenzhen 518055, Guangdong Province, China; Guangdong Cardiovascular Institute, Guangdong Provincial People's Hospital, Guangdong Academy of Medical Sciences, Guangzhou 510080, China.
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The mammalian genome contains thousands of regulatory elements whose activities are highly sensitive to external and internal stimulation. Here, we present a protocol to assess the murine cardiac function of regulatory elements using an adeno-associated virus serotype 9 (AAV9)-based massively parallel reporter assay (MPRA). We describe steps for the selection principle of enhancer candidates, MPRA library construction, animal delivery, sequencing library generations, and data analysis pipeline. Our MPRA protocol provides a comprehensive workflow for enhancer activity quantification in vivo. For complete details on the use and execution of this protocol, please refer to Yangpo Cao et al.1.
