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Genetics

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JoVE Journal Genetics
Stable Knockdown of Genes Encoding Extracellular Matrix Proteins in the C2C12 Myoblast Cell Line Using Small-Hairpin (sh)RNA
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Stable Knockdown of Genes Encoding Extracellular Matrix Proteins in the C2C12 Myoblast Cell Line Using Small-Hairpin (sh)RNA

Article DOI: 10.3791/60824-v • 12:19 min • February 12th, 2020
February 12th, 2020 •
1, 1, 1
1Orthopaedic Research Laboratories, Leni & Peter W. May Department of Orthopaedics, Icahn School of Medicine at Mount Sinai

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Summary
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We provide a protocol to stably knock down genes encoding extracellular matrix (ECM) proteins in C2C12 myoblasts using small-hairpin (sh) RNA. Targeting ADAMTSL2 as an example, we describe the methods for the validation of the knockdown efficiency on the mRNA, protein, and cellular level during C2C12 myoblast to myotube differentiation.

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Stable Knockdown Genes Extracellular Matrix Proteins C2C12 Myoblast Cell Line Small-hairpin RNA (shRNA) Function Of Proteins Myotube Formation Maturation Gene Knockdown Specific ShRNAs Analytical Tools Phenotyping C2C12 Cells Mature Myotubes Undifferentiated State Low Cell Density Routine Cell Culture Transfection Sodium Chloride PEI Stock Solution Plasmid DNA
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