Hiroki Kojima, Akiko Kashiwagi and Takashi Ikegami
Original Abstract: We performed single-cell RNA sequencing (scRNA-seq) on a population of 5,000 Tetrahymena thermophila, using the 10x Genomics 3′ gene expression analysis, to investigate gene expression variability within this clonal population. Initially, we estimated the 3ʹ-untranslated regions (3′ UTRs), which were absent in existing annotation files but are crucial for the 10x Genomics 3′ gene expression analysis, using the peaks2utr method. This allowed us to create a modified annotation file, which was then utilized in our scRNA-seq analysis. Our analysis revealed significant gene expression variability within the population, even after removing the effect of cell phase-related features. This variability predominantly appeared in six distinct clusters. Through gene ontology and KEGG pathway enrichment analyses, we identified that these were primarily associated with ribosomal proteins, proteins specific to mitochondria, proteins involved in peroxisome-specific carbon metabolism, cytoskeletal proteins, motor proteins, and immobilized antigens.
Summary (EN): Single-cell RNA sequencing of 5,000 clonal Tetrahymena thermophila cells (10x Genomics 3′), using the peaks2utr method to recover missing 3′ UTRs. Significant expression variability persisted after regressing out cell-phase effects, forming about six clusters tied to ribosomal, mitochondrial, peroxisomal, and cytoskeletal proteins—evidence of emerging sub-populations within a clonal population.
概要（日本語）: クローン集団であるテトラヒメナ5,000細胞に対し10x Genomicsの3′ scRNA-seqを実施。既存アノテーションに欠けていた3′ UTRをpeaks2utrで推定して解析した結果、細胞周期の影響を除いても顕著な遺伝子発現のばらつきが残り、約6つのクラスタを形成。リボソーム・ミトコンドリア・ペルオキシソーム・細胞骨格関連タンパク質と関連し、集団内に創発するサブ集団の存在を示唆する。
