Journal
STAR PROTOCOLS
Volume 2, Issue 3, Pages -Publisher
ELSEVIER
DOI: 10.1016/j.xpro.2021.100694
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Funding
- Association Francaise contre les Myopathies (AFM) [ANR-16-CE14-0032-01, AFM21711]
- LabexREVIVE [22946]
- Agence Nationale pour la Recherche (ANR) [ANR-10-LABX-73]
- Satnet [ANR-13-BSV1-0011-02]
- BMP-MyoStem [ANR-15-CE13-0011-01]
- MyoStemVasc [ANR-16-CE14-0002-03]
- muscleXTRA [ANR-17-CE14-0018-01]
- Fondation pour la Recherche Medicale (FRM) [ANR-19-CE13-0010]
- RHU CARMMA [ECO201806006793]
- [ANR-15-RHUS-0003]
- Agence Nationale de la Recherche (ANR) [ANR-19-CE13-0010, ANR-16-CE14-0032, ANR-17-CE14-0018] Funding Source: Agence Nationale de la Recherche (ANR)
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The single-nucleus RNA sequencing protocol optimized for adult mouse skeletal muscles allows sequencing of myonuclei in multinucleated myofibers and avoids transcriptional modifications induced by cell dissociation.
Single-nucleus RNA sequencing allows the profiling of gene expression in iso-lated nuclei. Here, we describe a step-by-step protocol optimized for adult mouse skeletal muscles. This protocol provides two main advantages compared to the widely used single-cell protocol. First, it allows us to sequence the myonu-clei of the multinucleated myofibers. Second, it circumvents the cell-dissociation -induced transcriptional modifications.For complete details on the use and execution of this protocol, please refer to Dos Santos et al. (2020) and Machado, Geara et al. (2021).
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