User:Vivek Rai/Mosaic-seq

Mosaic-seq (Mosaic single-cell analysis by indexed CRISPR sequencing) is a next-generation sequencing based method for high-throughput functional screening of enhancer sequences at a single-cell resolution.

Enhancers are a class of regulatory elements inside a cell that increase the expression of a target gene through transcription factor binding. They are known to orchestrate highly cell-type specific gene expression programs. Large scale consortium studies have predicted more than 2 million enhancer elements, although only a fraction of them are known to be functionally active. Thus, predicting whether an enhancer is active or not has important consequences. However, traditional methods such as reporter assays are too time-consuming to screen a large number of such elements.

Mosaic-seq uses a CRISPR-dCas9-KRAB based system to suppress target enhancer activity using a huge library of guide RNA. The targeted cells are then sequenced using single-cell RNA sequencing platform (Drop-seq) to measure the changes in transcriptome.