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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
 

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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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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Keywords: C2C12 Myoblast ShRNA Gene Knockdown Extracellular Matrix Proteins Stable Cell Line Transfection Puromycin Selection Cell Culture Myotube Formation
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