November 2024, Allshire Lab, Springer Nature Authors Dewornu, F.S., Tong, P., Torres-Garcia, S., Pidoux, A., Allshire, R., and Shukla, M. Summary By Dipika Mishra, Akiyoshi LabChromatin immunoprecipitation followed by sequencing (ChIP-Seq) is one of the most widely used tools in molecular biology to probe protein DNA interactions and identify histone modifications across the genome. The basic protocol involves crosslinking proteins to DNA using formaldehyde followed by fragmentation of cross-linked DNA. In the next step, the DNA interacting protein is immunoprecipitated using specific antibody, the crosslinks are reversed by heat and the associated DNA is then analyzed. However, there are certain limitations involving traditional ChIP-Seq. These include the time involved, the huge amount of sample needed and the high background noise generated. Researchers in Allshire lab have overcome these limitations by adapting ChIPmentation technology that integrates ChIP with transposase based tagmentation for fission yeast. Their protocol uses a hyperactive Tn5 transposase that enables both DNA fragmentation and adapter tagging or tagmentation of immunoprecipitated chromatin immobilized on the beads. The DNA recovered after tagmentation process is then subjected to amplification by PCR to add sequences required for next generation sequencing. This method thus provides a rapid, low input ChIP-Seq workflow and generates a higher resolution mapping of the fission yeast genome. Schematic overview of ChIPmentation workflow in fission yeast. Cells are cross-linked, followed by lysis and immunoprecipitation. Tagmentation of enriched chromatin is performed using Tn5 transposase. Tagmented DNA is recovered and amplified for library preparation and subsequent sequencing. Related Links Journal URL PI Website DOI This article was published on 2025-06-13
