Rapid and Low-Input Profiling of Histone Marks in Plants Using Nucleus CUT&Tag

Rapid and Low-Input Profiling of Histone Marks in Plants Using Nucleus CUT&Tag

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Characterizing genome-wide histone posttranscriptional modifications and transcriptional factor occupancy is crucial for deciphering their biological functions.Chromatin immunoprecipitation followed by sequencing (ChIP-seq) is a powerful method for genome-wide profiling of histone modifications and transcriptional factor-binding sites.However, the current ChIP-seq experimental procedure in plants requires significant material and several days for completion.CUT&Tag is an alternative method of ChIP-seq for low-sample and single-cell epigenomic Toners profiling using protein A-Tn5 transposase fusion proteins (PAT).In this study, we developed a nucleus CUT&Tag (nCUT&Tag) protocol based on the live-cell CUT&Tag technology.

Our results indicate that nCUT&Tag could be used for histone modifications profiling in both monocot rice and dicot rapeseed using crosslinked or fresh tissues.In addition, Clothing - Womens Tops - Hoodies both active and repressive histone marks such as H3K4me3 and H3K9me2 can be identified using our nCUT&Tag.More importantly, all the steps in nCUT&Tag can be finished in only 1 day, and the assay can be performed with as little as 0.01 g of plant tissue as starting materials.Therefore, our results demonstrate that nCUT&Tag is an efficient alternative strategy for plant epigenomic studies.