| Abstract [eng] |
Open chromatin analysis uses Mnase-seq, Chip-seq, and DNase-seq methods, which can detect epigenetic changes and provide information on transcription factor binding sites, chromatin availability at promoters, enhancers, or other regulatory elements. However, these methods require several million cells, thus losing the ability to study cells in rare populations. Assay for Transposase-Accessible Chromatin using sequencing or ATAC-seq is a method that can simultaneously fragment genomic DNA and attach sequencing adapters. This reduces the number of cells required from a few million to tens of thousands. This method is used to study the availability of chromatin, to determine the binding sites of regulatory elements. ATAC-seq currently uses a hyperactive trasposase that performs tagmentation. However, there are other transposases with similar activity. One such is the MuA transposase belonging to the same RNase H family. Therefore the aim of this work was to investigate whether MuA transposase can be used for chromatin tagmentation and ATAC-seq library preparation. This work demonstrates that the MuA transpososome, like the Tn5 transpososome, can tagment the nuclei of a selected human cell line. The optimal number of cells required for tagmentation with MuA transposase was determined. ATAC-seq libraries were prepared using MuA transposase and several different transposons that may provide specific advantages. Libraries were validated by sequencing showing that MuA transpososome can be used to analyze accessible chromatin. |
