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Single-cell RNA sequencing of meniscus from ACTA2-lineage traced mice, rats and dogs

ABSTRACT: 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.

INSTRUMENT(S): Biological safety cabinet, Centrifuge, Cell strainer, Automated cell counter/Hemocytometer, Surgical dissection tools, BD Rhapsody system, Computational server/cluster, Software: R (v4.3.x), Seurat package (v5.1.0), Illumina NovaSeq X, Microsyringe for intraperitoneal injection

ORGANISM(S): Rattus norvegicus

SUBMITTER: Mengyuan Zhang

PROVIDER: E-MTAB-16550 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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