Members of the same histone family that have diverged with differences in amino acid sequence, expression pattern, or chromatin localization are called histone variants. Histone variants and histone post-translational modifications are involved in epigenetic regulation and greatly enhance chromatin diversity. In plants, histone variants are involved in developmental processes and regulate plant responses to the environment. Lifeasible can help you study histone and variant modifications through chromatin immunoprecipitation sequencing (ChIP-seq) and cleavage under targets and tagmentation (CUT&Tag) related technologies.

CUT&Tag

With CUT&Tag, we can help you study the interaction of histones and transcription factors with DNA. We capture DNA-protein interactions by targeting the tag with an antibody fused to the Tn5 translocase, adding sequencing junctions to the target sequences in the process, and then conducting various analyses by purifying and sequencing the DNA fragments. This method is simple, easy to use, has a high signal-to-noise ratio, reproducible, requires as few as 60 cells, and promises to make ChIP-seq possible at the single-cell level.

ChIP-seq

We combine ChIP and NGS to isolate target modifications of proteins and their bound genomic DNA by antibody immunoprecipitation. We fragment and sequence the relevant DNA to determine the location and abundance of histone modifications on the genome. The process involves cell/tissue cross-linking, nucleus extraction, chromatin fragmentation, immunoprecipitation, DNA recovery, library construction, high-throughput sequencing, and bioinformatics analysis.

Plant species we have successfully analyzed

Varieties		Tissues
Oryza sativa Triticum aestivum L. Arabidopsis thaliana Solanum lycopersicum Nicotiana tabacum L. Brassica napus Brassica rapa subsp. pekinensis Vitis vinifera	Gossypium herbaceum Linn. Salvia miltiorrhiza Medicago sativa L. Malus domestica Glycine max Brachypodium sylvaticum Vernicia fordii Catharanthus roseus	Leaf blades, seedlings, shoots, stalks, rice spikes, healing tissue, etc.

Analysis of histone variants

We support the study of histone variants in vivo and in vitro using a variety of techniques, including knockdown studies of genes encoding specific variants (e.g., using RNAi knockdown), Co-IP, stable isotope labeling of amino acids, and quantitative mass spectrometry proteomics, immunohistochemistry, and protein blotting. The structure of nucleosomes can be altered by post-transcriptional modification (PTM) of histones and by including variant histones. We can analyze mono- and oligo-nucleosomes by enzyme-linked immunosorbent assay (ELISA). In addition, we can help you detect nucleosomes containing histone variants.

Lifeasible is committed to a variety of techniques to help you understand histones and their variant modifications and to elucidate how they function, which is critical for research in areas such as plant stress tolerance and development. Please feel free to contact us to submit your plant species of interest, and we will provide you with a customized solution.

*If your organization requires the signing of a confidentiality agreement, please contact us by email.

For research use only, not intended for any clinical use.

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