Probing cell identity hierarchies by fate titration and collision during direct reprogramming [scRNA-seq & scATAC-seq]. Probing cell identity hierarchies by fate titration and collision during direct reprogramming [scRNA-seq & scATAC-seq]

Collide-seq: A systematic comparison of how different reprogramming transcription factors achieve conversion as well as an in-depth analysis of how cells resolve the conflict when more than one fate is instructed. Overall design: Mouse embryonic fibroblasts were nucleofected with doxycycline inducible constructs for Ascl1, MyoD1 and a DNA binding mutant of Ascl1 (mutAscl1). Cells were expanded for about 10 days and positive cells sorted using FACS. All conditions were pooled and plated together. Transcription factor expression was induced for 72h via the administration of doxycycline. After 72h, cells were collected for scMultiome, i.e., scRNA-seq and scATAC-seq from the same cells

Collide-seq：本数据集旨在系统性对比不同重编程转录因子介导细胞命运转化的分子机制，并深入解析当细胞接收到多种命运指令时，其如何化解命运冲突的问题。
实验设计：将携带可经多西环素（doxycycline）诱导表达的Ascl1、MyoD1以及Ascl1的DNA结合突变体（mutAscl1）的重组载体，通过核转染技术导入小鼠胚胎成纤维细胞。将细胞扩增约10天后，利用荧光激活细胞分选术（FACS）分选阳性细胞。将所有实验组的细胞混合后共同铺板接种，通过添加多西环素诱导转录因子表达72小时。诱导72小时后，收集细胞进行单细胞多组学（scMultiome）测序，即对同一细胞同时开展单细胞RNA测序（scRNA-seq）与单细胞转座酶可及性测序（scATAC-seq）。