scRNA-Seq for RUNX1: replicate 1, nodox, run 2
We measured the impact of 115 RUNX1 Runt domain missense mutations in single myelogenous leukemia cells (K562) using the 10X Genomics Chromium platform. The library was generated from a modified lentiviral vector to contain the RUNX1 variant (WT, mutated, or GFP as LOF control) and a 12 base pair barcode sequence unique to each variant for identification during single cell transcriptome sequencing. The cells were transduced with the pooled variant library at a MOI of ~0.3 to ensure that each cell received a single construct. They were split into two populations, one treated with doxycycline to induce repression of the endogenous RUNX1 (dox), the other not (nodox). At day 7 post transduction, single cell RNA libraries were prepared and sequenced.