Division of Life Science, Center for Stem Cell Research, HKUST-Nan Fung Life Sciences Joint Laboratory, State Key Laboratory of Molecular Neuroscience, Molecular Neuroscience Center, The Hong Kong University of Science and Technology, Hong Kong, China.
Division of Life Science, Center for Stem Cell Research, HKUST-Nan Fung Life Sciences Joint Laboratory, State Key Laboratory of Molecular Neuroscience, Molecular Neuroscience Center, The Hong Kong University of Science and Technology, Hong Kong, China.
STAR Protoc. 2023 Sep 15;4(3):102376. doi: 10.1016/j.xpro.2023.102376. Epub 2023 Jun 22.
Chromatin accessibility is critical for cell identity. Conventional ATAC-seq can examine chromatin accessibility on freshly prepared muscle stem cells or satellite cells (SCs); however, isolating SCs in mice remains challenging. Here, we present a protocol to preserve the in vivo chromatin profile of SCs by applying paraformaldehyde (PFA) perfusion throughout the mouse before SC isolation. We describe steps for PFA perfusion and FACS sorting of SCs. We then detail library preparation for ATAC-seq. For complete details on the use and execution of this protocol, please refer to Dong et al..
染色质可及性对于细胞身份至关重要。传统的 ATAC-seq 可用于检测新鲜制备的肌肉干细胞或卫星细胞（SCs）中的染色质可及性；然而，在小鼠中分离SCs 仍然具有挑战性。在这里，我们提出了一种通过在 SC 分离前对小鼠进行多聚甲醛（PFA）灌注来保存 SC 体内染色质特征的方案。我们描述了 PFA 灌注和 SCs 的 FACS 分选步骤。然后，我们详细介绍了 ATAC-seq 的文库制备。有关此方案的使用和执行的完整详细信息，请参阅 Dong 等人的研究。
