Project description:Chromatin endogenous cleavage (ChEC) uses fusion of a protein of interest to micrococcal nuclease (MNase) to target calcium-dependent cleavage to specific genomic loci in vivo. Here we report the combination of ChEC with high-throughput sequencing (ChEC-seq) to map budding yeast transcription factor (TF) binding. Temporal analysis of ChEC-seq data reveals two classes of sites for TFs, one displaying rapid cleavage at sites with robust consensus motifs and the second showing slow cleavage at largely unique sites with low-scoring motifs. Sites with high-scoring motifs also display asymmetric cleavage, indicating that ChEC-seq provides information on the directionality of TF-DNA interactions. Strikingly, similar DNA shape patterns are observed regardless of motif strength, indicating that the kinetics of ChEC-seq discriminates DNA recognition through sequence and/or shape. We propose that time-resolved ChEC-seq detects both high-affinity interactions of TFs with consensus motifs and sites preferentially sampled by TFs during diffusion and sliding.
Project description:We optimized the SHAPE-MaP (selective 2'-hydroxyl acylation analyzed by primer extension and mutational profiling) assay to analyze structures of paired circular (circSHAPE-MaP) and linear (linearSHAPE-MaP) RNAs.
Project description:We combined ChIP-seq of chromatin marks and key islet transcription factor with RNA-seq in human islets to map cis-regulatory networks in this primary tissue. The output of this project provides a reference map to dissect genetic variants that alter the susceptibility for Type 2 diabetes, and assist efforts to generate new beta-cells by transcriptional programming strategies
Project description:To investigate the gene expression in dendritic cells after treating hemiphilic mice with HBSS, plasma derived or recombinant factor VIII Gene expression in dendritic cells were measured 24 hours post treatment
