Project description:A library of cassette exon splicing reporter mini-genes was cloned under the control of five different promoters and tested, alongside pharmacological treatments affecting transcription, by transfection into HEK293 cells, isolation of the mRNA and targeted high-throughput sequencing of the reporter constructs to determine the effect of promoter identity and pharmacological treatment on splicing behavior.
Project description:Interactions among cis-regulatory elements (CREs) are central to mammalian gene regulation. Long @$$ massively parallel reporter assays (LAMPRAs) integrate combinatorial cloning, molecular barcoding and long- and short-read sequencing, to scalably measure how CRE identities, numbers, spacings, orders, orientations, and interactions shape regulatory output at multi-kilobase length scales. As a proof-of-concept, we assay 36,000 x 5-kb synthetic cis-regulatory loci (sCRLs), each a 5 x 1-kb random combination of enhancers, insulators and spacers, to model how locus composition drives gene expression.