Immunology Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC 3052, Australia; Epigenetics and Development Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC 3052, Australia; Department of Medical Biology, University of Melbourne, Parkville, VIC 3052, Australia.
Immunology Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC 3052, Australia; Department of Medical Biology, University of Melbourne, Parkville, VIC 3052, Australia.
Immunity. 2021 Jun 8;54(6):1338-1351.e9. doi: 10.1016/j.immuni.2021.03.012. Epub 2021 Apr 15.
Despite advances in single-cell multi-omics, a single stem or progenitor cell can only be tested once. We developed clonal multi-omics, in which daughters of a clone act as surrogates of the founder, thereby allowing multiple independent assays per clone. With SIS-seq, clonal siblings in parallel "sister" assays are examined either for gene expression by RNA sequencing (RNA-seq) or for fate in culture. We identified, and then validated using CRISPR, genes that controlled fate bias for different dendritic cell (DC) subtypes. This included Bcor as a suppressor of plasmacytoid DC (pDC) and conventional DC type 2 (cDC2) numbers during Flt3 ligand-mediated emergency DC development. We then developed SIS-skew to examine development of wild-type and Bcor-deficient siblings of the same clone in parallel. We found Bcor restricted clonal expansion, especially for cDC2s, and suppressed clonal fate potential, especially for pDCs. Therefore, SIS-seq and SIS-skew can reveal the molecular and cellular mechanisms governing clonal fate.
尽管单细胞多组学技术取得了进展，但单个干细胞或祖细胞只能被检测一次。我们开发了克隆多组学技术，其中克隆的后代可以作为创始人的替代品，从而允许每个克隆进行多个独立的检测。通过 SIS-seq，可以同时对平行的“姐妹”克隆分析进行基因表达的 RNA 测序（RNA-seq）或培养中的命运分析。我们确定了控制不同树突状细胞 (DC) 亚型命运偏向的基因，然后使用 CRISPR 进行了验证。这包括 Bcor 作为 Flt3 配体介导的应急 DC 发育过程中浆细胞样 DC (pDC) 和传统 DC 类型 2 (cDC2) 数量的抑制因子。然后，我们开发了 SIS-skew 来同时平行检测同一克隆的野生型和 Bcor 缺陷型兄弟姐妹的发育情况。我们发现 Bcor 限制了克隆的扩增，尤其是 cDC2，并且抑制了克隆的命运潜力，尤其是 pDC。因此，SIS-seq 和 SIS-skew 可以揭示控制克隆命运的分子和细胞机制。
