Published January 2, 2025 | Version v2

Optimized Analytical Workflow for Single-Nucleus Transcriptomics in Main Metabolic Tissues

  • 1. Shanghai Key Laboratory of Metabolic Remodelling and Health, Institute of Metabolism and Integrative Biology, Centre for Evolutionary Biology, Fudan University, Shanghai, 200438, China

Description

Single-nucleus RNA sequencing (snRNA-seq) has emerged as a powerful approach for studying cellular heterogeneity in metabolic tissues. However, snRNA-seq analysis remains challenging due to low gene expression and data complexity. Here, we introduce an optimized analytical workflow for snRNA-seq data from 67 samples across four main metabolic tissues white adipose tissue, hypothalamus, muscle and liver. We emphasized the importance of key steps including ambient RNA removal, doublet identification, normalization and data integration to ensure accurate downstream analysis. This workflow offers a valuable resource for researchers in metabolism, facilitating deeper insights into cellular diversity and metabolic function through rigorous snRNA-seq analysis.

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Additional details

References

  • Scripture-Adams, D.D., et al., Single nuclei transcriptomics of muscle reveals intra-muscular cell dynamics linked to dystrophin loss and rescue. Communications Biology, 2022. 5(1): p. 989.
  • Massier, L., et al., An integrated single cell and spatial transcriptomic map of human white adipose tissue. Nature Communications, 2023. 14(1): p. 1438.
  • Gribben, C., et al., Acquisition of epithelial plasticity in human chronic liver disease. Nature, 2024. 630(8015): p. 166-173.
  • Huang, Y., et al., Maternal dietary fat during lactation shapes single nucleus transcriptomic profile of postnatal offspring hypothalamus in a sexually dimorphic manner in mice. Nature Communications, 2024. 15(1): p. 2382.