Single-cell RNA sequencing of meniscus from ACTA2-lineage traced mice, rats and dogs
This study presents a cross-species single-cell transcriptomic atlas of the meniscus. The analysis includes genetic lineage tracing mice, wild-type rats, and dogs. Genetic Lineage Tracing Mice: ACTA2-CreERT mice (generated via CRISPR/Cas9 knock-in at the Acta2 locus) were crossed with R26-CAG-LSL-tdTomato reporter mice. All mice received tamoxifen (75 mg/kg, i.p. for 5 days) to induce tdTomato expression in ACTA2-lineage cells. Menisci were harvested from an early post-induction cohort at 2 weeks (n=13) and a late cohort at 10 weeks of age (n=10). Other Species: Menisci were also analyzed from 12-week-old male Sprague-Dawley rats (n=4) and 12-month-old male beagles (n=2). Single-cell suspensions were captured and barcoded using the BD Rhapsody system. Libraries were constructed with the BD Resolve Whole-Transcriptome Amplification workflow and sequenced on Illumina platforms. Bioinformatic analysis was performed using Seurat for integration, clustering, and annotation of cell types, followed by subpopulation analysis and trajectory inference.
This study presents a cross-species single-cell transcriptomic atlas of the meniscus. The analysis includes genetic lineage tracing mice, wild-type rats, and dogs. Genetic Lineage Tracing Mice: ACTA2-CreERT mice (generated via CRISPR/Cas9 knock-in at the Acta2 locus) were crossed with R26-CAG-LSL-tdTomato reporter mice. All mice received tamoxifen (75 mg/kg, i.p. for 5 days) to induce tdTomato expression in ACTA2-lineage cells. Menisci were harvested from an early post-induction cohort at 2 weeks (n=13) and a late cohort at 10 weeks of age (n=10). Other Species: Menisci were also analyzed from 12-week-old male Sprague-Dawley rats (n=4) and 12-month-old male beagles (n=2). Single-cell suspensions were captured and barcoded using the BD Rhapsody system. Libraries were constructed with the BD Resolve Whole-Transcriptome Amplification workflow and sequenced on Illumina platforms. Bioinformatic analysis was performed using Seurat for integration, clustering, and annotation of cell types, followed by subpopulation analysis and trajectory inference.
European Bioinformatics Institute