Single_cell_RNA_seq_analysis_of_CRISPR_mutagenised_cells

In this study we aim to develop a novel method that allows us to analyse transcriptome changes after gene perturbation at a scale. To this end, we employ the CRISPR-Cas9 and single-cell RNA-sequencing technologies. Cas9-expressing cells will be first transduced with a pool of lentiviruses, each of which carries a gRNA targeting a gene and an index together with a fluorescent reporter gene. Transduction will be conducted at a low MOI, so that each of transduced cells is likely to receive one viral particle. At an appropriate time point, the transduced cells will be collected by cell sorting and subjected to single-cell RNA-seq library construction and resulting libraries will be sequenced on an Illumina platform.