Expression profiling by high throughput sequencing
Firstly, we extracted bone marrow cells from AML mouse models in four different timepoints (T0, T1, T2 and T3). And we used droplet-based single cell RNA sequencing technology to reveal the early-MDS, late-MDS, MDS-AML and AML four-stage landscape in a mouse AML model initiated by Myc overexpression. Also, we extracted one AML patient bone marrow sample to sequence by constructing library following 10X genomics platform. And integrated this data with four AML patients data from 10X genomics database to explain the molecular mechanism of splicing events of TMEM134 in AML. To verify the alternative splicing results, we used smartseq2 to construct the single cell library in T3. And combined TCGA-LAML and TARGET-AML database and our mouse function data, we identify the alternative splicing events of TMEM134 could drive the proliferation function in acute myelogenous leukemogenesis.
Single cell RNA sequencing (scRNA-seq) in leukemogenesis