Name: Reusable Single Cell for Iterative Epigenomic Analyses (Video) | JoVE Journal
Uploaded: 2022-02-11T13:30:00.000+00:00
Duration: 10 min 28 s
Description: Learn reusable single-cell technology for iterative epigenomic analyses using the same cell multiple times. This method enables analysis of multiple epigenetic marks and transcription factors with statistical validation.

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5 minutes

We thank Drs. David Sanchez-Martin and Christopher B. Buck for comments during the conceptualization stage of the project. We also thank the Genomics Core, Center for Cancer Research, National Cancer Institute, National Institutes of Health for help in preliminary experiments, and the Collaborative Bioinformatics Resource, CCR, NCI, NIH for advice in computational analysis. We thank Ms. Anna Word for helping with the optimization of DNA polymerases used in the method. This work utilized the computational resources of the NIHHPC Biowulf cluster (http://hpc.nih.gov). This project is supported by the Intramural Program of the Center for Cancer Research, National Cancer Institute, National Institutes of Health, the NCI Director&#39;s Innovation Award (#397172), and Federal funds from the National Cancer Institute under Contract No. HHSN261200800001E. We thank Drs. Tom Misteli, Carol Thiele, Douglas R. Lowy, and all members of Laboratory of Cellular Oncology for productive comments.&#160;

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Drs. Ohnuki and Tosato are co-inventors on a patent entitled "Methods for preparing a reusable single cell and methods for analyzing the epigenome, transcriptome and genome of a single cell" (EP3619307 and US20200102604). The patent application was filed in part based on preliminary results related to the technology described in the current manuscript. The invention or inventions described and claimed in this patent application were made while the inventors were full-time employees of the U.S. Government. Therefore, under 45 Code of Federal Regulations Part 7, all rights, title, and interest to this patent application have been or should by law be assigned to the U.S. Government. The U.S. Government conveys a portion of the royalties it receives to its employee inventors under 15 U.S. Code &#167; 3710c.

This article describes the step-by-step protocol for the recently reported single-cell multiepigenomic analysis using reusable single cells7. In the subsequent paragraphs, we discuss critical points, emphasizing potential limitations in the protocol. 
One of the critical points throughout the protocol (from steps 7.2-13) is avoiding DNase contamination. A single cell only has two copies of genomic DNA. Therefore, damaging genomic DNA critically reduces the signal number...

Single-cell technology is entering the era of single-cell multiomics, which integrates individual single-cell omics technologies1. Recently, single-cell transcriptomics has been combined with methods for detecting chromatin accessibility (scNMT-seq2 and SHARE-seq3) or histone modifications (Paired-seq4 and Paired-Tag5). More recently, single-cell transcriptomics and proteomics were integrated with chromatin accessibility (DOGMA-seq6). These methods use transposase-based tagging for detecting chromatin accessibility or histone modifications.
Transposase-based approaches cleave genomic DNA and add a DNA barcode at the end of the genomic DNA fragment. Each cleaved genomic fragment can only accept up to two DNA barcodes (= one epigenetic mark per cleavage site), and the genomic DNA at the cleavage site is lost. Therefore, cleavage-based approaches have a trade-off between the number of epigenetic marks tested and the signal density. This hampers the analysis of multiple epigenetic marks in the same single cell. A single-cell epigenomic method that does not cleave the genomic DNA was developed to overcome this issue7,8.
In addition to the cleavage-derived issue mentioned above, transposase-based approaches have other limitations. In single-cell epigenome analysis, it is critical to know the location of histones and DNA-associated proteins on the genome. In current approaches, this is accomplished by using unfixed single cells and retention of protein-DNA and protein-protein interactions. However, this generates strong bias to accessible chromatin regions, even in the analysis of histone modifications 9. The location of histones and genome-associated proteins on the genome can be preserved without crosslinking protein-DNA and protein-protein, using a polyacrylamide scaffold7,8. This approach reduces the structural bias observed in current approaches that depend upon protein-DNA and protein-protein interactions.
Transposase-based approaches can acquire signals only once from a single cell. Therefore, it is difficult to delineate the complete epigenome of a single cell due to the drop-off of the signals. Reusable single cells have been developed to overcome current limitations by allowing iterative epigenomic analysis in the same single cell.

<table><tbody><tr><td>10x CutSmart buffer</td><td>New England BioLabs</td><td>B6004</td><td>10x Digestion buffer</td></tr><tr><td>200 proof ethanol</td><td>Warner-Graham Company</td><td>200 proof</td><td>Ethanol</td></tr><tr><td>5-Hydroxymethylcytosine (5-hmC) Monoclonal Antibody [HMC/4D9]</td><td>Epigentek</td><td>A-1018-100</td><td>Anti-5hmC</td></tr><tr><td>Acridine Orange/Propidium Iodide Stain</td><td>Logos Biosystems</td><td>F23001</td><td>Cell counter</td></tr><tr><td>Acrylamide solution, 40% in H2O, for molecular biology</td><td>MilliporeSigma</td><td>01697-500ML</td><td>40% acrylamide solution</td></tr><tr><td>All-in-One Fluorescence Microscope BZ-X710</td><td>Keyence</td><td>BZ-X710</td><td>Scanning microscope</td></tr><tr><td>Amicon Ultra-0.5 Centrifugal Filter Unit</td><td>MilliporeSigma</td><td>UFC510024</td><td>Ultrafiltration cassette</td></tr><tr><td>Ammonium persulfate for molecular biology</td><td>MilliporeSigma</td><td>A3678-100G</td><td>Ammonium persulfate powder</td></tr><tr><td>Anhydrous DMF</td><td>Vector laboratories</td><td>S-4001-005</td><td>Anhydrous N,N-dimethylformamide (DMF)</td></tr><tr><td>Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8]</td><td>Abcam</td><td>ab5408</td><td>Anti-Pol II</td></tr><tr><td>Anti-TRAP220/MED1 (phospho T1457) antibody</td><td>Abcam</td><td>ab60950</td><td>Anti-Med1</td></tr><tr><td>BciVI</td><td>New England BioLabs</td><td>R0596L</td><td>BciVI</td></tr><tr><td>Bovine Serum Albumin solution, 20 mg/mL in H&lt;sub&gt;2&lt;/sub&gt;O, low bioburden, protease-free, for molecular biology</td><td>MilliporeSigma</td><td>B8667-5ML</td><td>20% BSA (Table 7)</td></tr><tr><td>Bst DNA Polymerase, Large Fragment</td><td>New England BioLabs</td><td>M0275L</td><td>Bst DNA polymerase</td></tr><tr><td>BT10 Series 10 &amp;micro;l Barrier Tip</td><td>NEPTUNE</td><td>BT10</td><td>P10 low-retention tip</td></tr><tr><td>CellCelector</td><td>Automated Lab Solutions</td><td>N/A</td><td>Automated single cell picking robot</td></tr><tr><td>CellCelector 4 nl nanowell plates for single cell cloning, Plate S200-100 100K, 24 well,ULA</td><td>Automated Lab Solutions</td><td>CC0079</td><td>4 nL nanowell plate</td></tr><tr><td>Chloroform</td><td>MilliporeSigma</td><td></td><td>Chloroform</td></tr><tr><td>Corning Costar 96-Well, Cell Culture-Treated, Flat-Bottom Microplate</td><td>Corning</td><td>3596</td><td>Flat-bottom 96-well plates</td></tr><tr><td>Deep Vent (exo-) DNA Polymerase</td><td>New England BioLabs</td><td>M0259L</td><td>Exo&lt;sup&gt;-&lt;/sup&gt; DNA polymerase</td></tr><tr><td>DNA LoBind Tubes, 0.5 mL</td><td>Eppendorf</td><td>30108035</td><td>0.5 mL DNA low-binding tube</td></tr><tr><td>DNA Oligo, 1st random primer</td><td>Integrated DNA Technologies</td><td>N/A, see Table 3</td><td>1st random primer</td></tr><tr><td>DNA Oligo, 2nd random primer Cell#01</td><td>Integrated DNA Technologies</td><td>N/A, see Table 3</td><td>2nd random primer</td></tr><tr><td>DNA Oligo, 2nd random primer Cell#02</td><td>Integrated DNA Technologies</td><td>N/A, see Table 3</td><td>2nd random primer</td></tr><tr><td>DNA Oligo, 2nd random primer Cell#03</td><td>Integrated DNA Technologies</td><td>N/A, see Table 3</td><td>2nd random primer</td></tr><tr><td>DNA Oligo, 2nd random primer Cell#04</td><td>Integrated DNA Technologies</td><td>N/A, see Table 3</td><td>2nd random primer</td></tr><tr><td>DNA Oligo, 2nd random primer Cell#05</td><td>Integrated DNA Technologies</td><td>N/A, see Table 3</td><td>2nd random primer</td></tr><tr><td>DNA Oligo, 2nd random primer Cell#06</td><td>Integrated DNA Technologies</td><td>N/A, see Table 3</td><td>2nd random primer</td></tr><tr><td>DNA Oligo, 2nd random primer Cell#07</td><td>Integrated DNA Technologies</td><td>N/A, see Table 3</td><td>2nd random primer</td></tr><tr><td>DNA Oligo, 2nd random primer Cell#08</td><td>Integrated DNA Technologies</td><td>N/A, see Table 3</td><td>2nd random primer</td></tr><tr><td>DNA Oligo, 2nd random primer Cell#09</td><td>Integrated DNA Technologies</td><td>N/A, see Table 3</td><td>2nd random primer</td></tr><tr><td>DNA Oligo, 2nd random primer Cell#10</td><td>Integrated DNA Technologies</td><td>N/A, see Table 3</td><td>2nd random primer</td></tr><tr><td>DNA Oligo, 2nd random primer Cell#11</td><td>Integrated DNA Technologies</td><td>N/A, see Table 3</td><td>2nd random primer</td></tr><tr><td>DNA Oligo, 2nd random primer Cell#12</td><td>Integrated DNA Technologies</td><td>N/A, see Table 3</td><td>2nd random primer</td></tr><tr><td>DNA Oligo, 2nd synthesis primer</td><td>Integrated DNA Technologies</td><td>N/A, see Table 3</td><td>2nd synthesis primer</td></tr><tr><td>DNA Oligo, Ligation Adaptor</td><td>Integrated DNA Technologies</td><td>N/A, see Table 3</td><td>Ligation Adaptor</td></tr><tr><td>DNA Oligo, Reverse Transcription primer</td><td>Integrated DNA Technologies</td><td>N/A, see Table 3</td><td>Reverse Transcription primer</td></tr><tr><td>DNase I (RNase-free)</td><td>New England BioLabs</td><td>M0303L</td><td>DNase I (RNase-free, 4 U).</td></tr><tr><td>DNase I Reaction Buffer</td><td>New England BioLabs</td><td>B0303S</td><td>10x DNase I buffer (NEB)</td></tr><tr><td>dNTP Mix (10 mM each)</td><td>Thermo Fisher</td><td>R0192</td><td>10 mM dNTPs</td></tr><tr><td>Fetal Bovine Serum, USA origin, Heat-inactivated</td><td>MilliporeSigma</td><td>F4135-500ML</td><td>Fetal bovine serum</td></tr><tr><td>HiScribe T7 High Yield RNA Synthesis Kit</td><td>New England BioLabs</td><td>E2040S</td><td>In-vitro-transcription master mix</td></tr><tr><td>Histone H3K27ac antibody</td><td>Active motif</td><td>39133</td><td>Anti-H3K27ac</td></tr><tr><td>Histone H3K27me3 antibody</td><td>Active motif</td><td>39155</td><td>Anti-H3K27me3</td></tr><tr><td>IgG from rabbit serum</td><td>Millipore Sigma</td><td>I5006-10MG</td><td>Control IgG</td></tr><tr><td>Iron oxide(II,III) magnetic nanopowder, 30 nm avg. part. size (TEM), NHS ester functionalized</td><td>MilliporeSigma</td><td>747467-1G</td><td>NHS ester functionalized 30 nm iron oxide powder</td></tr><tr><td>K-562</td><td>American Type Culture Collection (ATCC)</td><td>CCL-243</td><td>cells</td></tr><tr><td>Linear Acrylamide (5 mg/mL)</td><td>Thermo Fisher</td><td>AM9520</td><td>Linear Acrylamide</td></tr><tr><td>LUNA-FL Dual Fluorescence Cell Counter</td><td>Logos Biosystems</td><td>L20001</td><td>Cell counter</td></tr><tr><td>LUNA Cell Counting Slides, 50 Slides</td><td>Logos Biosystems</td><td>L12001</td><td>Cell counter</td></tr><tr><td>Mineral oil, BioReagent, for molecular biology, light oil</td><td>MilliporeSigma</td><td>M5904-500ML</td><td>Mineral oil</td></tr><tr><td>N,N,N&amp;prime;,N&amp;prime;-Tetramethylethylenediamine for molecular biology</td><td>MilliporeSigma</td><td>T7024-100ML</td><td>N,N,N&amp;prime;,N&amp;prime;-Tetramethylethylenediamine</td></tr><tr><td>NaCl (5 M), RNase-free</td><td>Thermo Fisher</td><td>AM9760G</td><td>5M NaCl</td></tr><tr><td>NanoDrop Lite</td><td>Thermo Fisher</td><td>2516</td><td>&amp;nbsp;Microvolume spectrophotometer</td></tr><tr><td>NEST 2 mL 96-Well Deep Well Plate, V Bottom</td><td>Opentrons</td><td>N/A</td><td>2 mL deep well 96-well plate</td></tr><tr><td>Non-skirted 96-well PCR plate</td><td>Genesee Scientific</td><td>27-405</td><td>96-well PCR plate</td></tr><tr><td>NuSive GTG Agarose</td><td>Lonza</td><td>50081</td><td>Agarose</td></tr><tr><td>OmniPur Acrylamide: Bis-acrylamide 19:1, 40% Solution</td><td>MilliporeSigma</td><td>1300-500ML</td><td>40%Acrylamide/Bis-acrylamide</td></tr><tr><td>OT-2 lab robot</td><td>Opentrons</td><td>OT2</td><td>Automated liquid handling robot</td></tr><tr><td>Paraformaldehyde, EM Grade, Purified, 20% Aqueous Solution</td><td>Electron Microscopy Sciences</td><td>15713</td><td>20% Pararmaldehyde</td></tr><tr><td>PBS (10x), pH 7.4</td><td>Thermo Fisher</td><td>70011044</td><td>10x PBS</td></tr><tr><td>PIPETMAN Classic P1000</td><td>GILSON</td><td>F123602</td><td>A P1000 pipette</td></tr><tr><td>Protein LoBind Tubes, 1.5 mL</td><td>Eppendorf</td><td>925000090</td><td>1.5 mL Protein low-binding tube</td></tr><tr><td>QIAgen Gel Extraction kit</td><td>Qiagen</td><td>28706</td><td>A P1000 pipette</td></tr><tr><td>Quant-iT PicoGreen dsDNA Assay</td><td>Thermo Fisher</td><td>P11495</td><td>&amp;nbsp;dsDNA specific intercalator dye</td></tr><tr><td>Quick Ligation kit</td><td>New England BioLabs</td><td>M2200L</td><td>T4 DNA ligase (NEB)</td></tr><tr><td>RNaseOUT Recombinant Ribonuclease Inhibitor</td><td>Thermo Fisher</td><td>10777019</td><td>RNAse inhibitor</td></tr><tr><td>S-4FB Crosslinker (DMF-soluble)</td><td>Vector laboratories</td><td>S-1004-105</td><td>Succinimidyl 4-formylbenzoate (S-4FB)</td></tr><tr><td>S-HyNic</td><td>Vector laboratories</td><td>S-1002-105</td><td>Succinimidyl 6-hydrazinonicotinate acetone hydrazone (S-HyNic)</td></tr><tr><td>Sodium Acetate, 3 M, pH 5.2, Molecular Biology Grade</td><td>MilliporeSigma</td><td>567422-100ML</td><td>3M Sodium acetate (pH 5.2)</td></tr><tr><td>Sodium bicarbonate, 1M buffer soln., pH 8.5</td><td>Alfa Aesar</td><td>J60408</td><td>1M sodium bicarbonate buffer, pH 8.5</td></tr><tr><td>Sodium phosphate dibasic for molecular biology</td><td>MilliporeSigma</td><td>S3264-250G</td><td>Na&lt;sub&gt;2&lt;/sub&gt;HPO4</td></tr><tr><td>Sodium phosphate monobasic for molecular biology</td><td>MilliporeSigma</td><td>S3139-250G</td><td>NaH&lt;sub&gt;2&lt;/sub&gt;PO4</td></tr><tr><td>SuperScript IV reverse transcriptase</td><td>Thermo Fisher</td><td>18090050</td><td>Reverse transcriptase</td></tr><tr><td>SYBR Gold Nucleic Acid Gel Stain (10,000x Concentrate in DMSO)</td><td>Thermo Fisher</td><td>S11494</td><td>An intercalator dye for DNA</td></tr><tr><td>T4 DNA Ligase Reaction Buffer</td><td>New England BioLabs</td><td>B0202S</td><td>10x T4 DNA ligase reaction buffer</td></tr><tr><td>ThermoPol Reaction Buffer Pack</td><td>New England BioLabs</td><td>B9004S</td><td>10x TPM-T buffer (Tris-HCl/Pottasium chloride/Magnesium sulfate/Triton X-100)</td></tr><tr><td>TRIzol LS reagent</td><td>Thermo Fisher</td><td>10296-028</td><td>Guanidinium thiocyanate-phenol-chloroform extraction</td></tr><tr><td>TruSeq Nano DNA library prep kit</td><td>Illumina</td><td>20015965</td><td>A DNA library preparation kit (see also the manufacturer&#039;s instruction)</td></tr><tr><td>Ultramer DNA Oligo, Anti-5hmC_Ab#005</td><td>Integrated DNA Technologies</td><td>N/A, see Table 3</td><td>An amine-modified DNA probe for antibody</td></tr><tr><td>Ultramer DNA Oligo, Anti-H3K27ac_Ab#002</td><td>Integrated DNA Technologies</td><td>N/A, see Table 3</td><td>An amine-modified DNA probe for antibody</td></tr><tr><td>Ultramer DNA Oligo, Anti-H3K27me3_Ab#003</td><td>Integrated DNA Technologies</td><td>N/A, see Table 3</td><td>An amine-modified DNA probe for antibody</td></tr><tr><td>Ultramer DNA Oligo, Anti-Med1_Ab#004</td><td>Integrated DNA Technologies</td><td>N/A, see Table 3</td><td>An amine-modified DNA probe for antibody</td></tr><tr><td>Ultramer DNA Oligo, Anti-Pol II_Ab#006</td><td>Integrated DNA Technologies</td><td>N/A, see Table 3</td><td>An amine-modified DNA probe for antibody</td></tr><tr><td>Ultramer DNA Oligo, Control IgG_Ab#001</td><td>Integrated DNA Technologies</td><td>N/A, see Table 3</td><td>An amine-modified DNA probe for control IgG</td></tr><tr><td>UltraPure 0.5 M EDTA, pH 8.0</td><td>Thermo Fisher</td><td>15575020</td><td>0.5M EDTA, pH 8.0</td></tr><tr><td>UltraPure DNase/RNase-Free Distilled Water</td><td>Thermo Fisher</td><td>10977023</td><td>Ultrapure water</td></tr><tr><td>Zeba Splin Desalting Columns, 7K MWCO, 0.5 mL</td><td>Thermo Fisher</td><td>89882</td><td>Desalting column</td></tr></tbody></table>

reusable single cell for iterative epigenomic analyses

Reusable Single Cell for Iterative Epigenomic Analyses

Current single-cell epigenome analyses are designed for single use. The cell is discarded after a single use, preventing analysis of multiple epigenetic marks in a single cell and requiring data from other cells to distinguish signal from experimental background noise in a single cell. This paper describes a method to reuse the same single cell for iterative epigenomic analyses. 
In this experimental method, cellular proteins are first anchored to a polyacrylamide polymer instead of crosslinking them to protein and DNA, alleviating structural bias. This critical step allows repeated experiments with the same single cell. Next, a random primer with a scaffold sequence for proximity ligation is annealed to the genomic DNA, and the genomic sequence is added to the primer by extension using a DNA polymerase. Subsequently, an antibody against an epigenetic marker and control IgG, each labeled with different DNA probes, are bound to the respective targets in the same single cell. 
Proximity ligation is induced between the random primer and the antibody by adding a connector DNA with complementary sequences to the scaffold sequence of the random primer and the antibody-DNA probe. This approach integrates antibody information and nearby genome sequences in a single DNA product of proximity ligation. By enabling repeated experiments with the same single cell, this method allows an increase in data density from a rare cell and statistical analysis using only IgG and antibody data from the same cell. The reusable single cells prepared by this method can be stored for at least a few months and reused later to broaden epigenetic characterization and increase data density. This method provides flexibility to researchers and their projects.

K562 single cells were generated using the protocol described in step 8 (see Figure 5). Single cells were embedded in the outer layer of the polyacrylamide bead. Cell DNA was stained and visualized using an intercalator dye for DNA staining.
<img alt="Microscopy image of microdroplets in liquid, marked for analysis in optical study." class="xfigimg" src="/files/ftp_upload/63456/63456fig05.jpg" data-altupdatedat="1788282455000" data...

The present protocol describes a single-cell method for iterative epigenomic analyses using a reusable single cell. The reusable single cell allows analyses of multiple epigenetic marks in the same single cell and statistical validation of the results.

Current single-cell epigenome analyses are designed for single use. The cell is discarded after a single use, preventing analysis of multiple epigenetic ...

reusable-single-cell-for-iterative-epigenomic-analyses

Research

JoVE Journal

Genetics

In this article we describe a simple method for the harvesting of single cells from rat primary neuronal cultures and subsequent transcriptome analysis using aRNA amplification. This approach is generalizable to any cell type.

Transcriptome Analysis of Single Cells

Methods for mapping in vivo protein-DNA interactions are becoming crucial for every aspect of genomic research but they are laborious, costly, and time consuming. Here a commercially available robotic liquid handling system that automates chromatin immunoprecipitation for mapping in vivo protein-DNA interactions with limited amounts of cells is presented.

Automating ChIP-seq Experiments to Generate Epigenetic Profiles on 10,000 HeLa Cells

Single-cell sequencing reveals genotypic heterogeneity in biological systems, but current technologies lack the throughput necessary for the deep profiling of community composition and function. Here, we describe a microfluidic workflow for sequencing &gt;50,000 single-cell genomes from diverse cell populations.

An Ultrahigh-throughput Microfluidic Platform for Single-cell Genome Sequencing

Bulk gene expression measurements cloud individual cell differences in heterogeneous cell populations. Here, we describe a protocol for how single-cell gene expression analysis and index sorting by Florescence Activated Cell Sorting (FACS) can be combined to delineate heterogeneity and immunophenotypically characterize molecularly distinct cell populations.

A Combinatorial Single-cell Approach to Characterize the Molecular and Immunophenotypic Heterogeneity of Human Stem and Progenitor Populations

This protocol describes the general processes and quality control checks necessary for preparing healthy adult mammalian single cells for droplet-based, high throughput single cell RNA-Seq preparations. Sequencing parameters, read alignment, and downstream single-cell bioinformatic analysis are also provided.

Droplet Barcoding-Based Single Cell Transcriptomics of Adult Mammalian Tissues

Here we presented a multiplexed single cell mRNA sequencing method to profile gene expression in mouse embryonic tissues. The droplet-based single cell mRNA sequencing (scRNA-Seq) method in combination with multiplexing strategies can profile single cells from multiple samples simultaneously, which significantly reduces reagent costs and minimizes experimental batch effects.

Multiplexed Single Cell mRNA Sequencing Analysis of Mouse Embryonic Cells

CUT&amp;RUN and its variants can be used to determine protein occupancy on chromatin. This protocol describes how to determine protein localization on chromatin using single-cell uliCUT&amp;RUN.

Single-Cell Factor Localization on Chromatin using Ultra-Low Input Cleavage Under Targets and Release using Nuclease

In this protocol, the translating ribosome affinity purification (TRAP) method and the isolation of nuclei tagged in specific cell types (INTACT) method were optimized for the paired interrogation of the cell-specific ovarian transcriptome and epigenome using the NuTRAP mouse model crossed to a Cyp17a1-Cre mouse line.

Cell-Specific Paired Interrogation of the Mouse Ovarian Epigenome and Transcriptome

The protocol combines human intestinal organoid technology with single cell transcriptomic analysis to provide significant insight into previously unexplored intestinal biology.

Using Human Intestinal Organoids to Understand the Small Intestine Epithelium at the Single Cell Transcriptional Level

This study presents a spatial Trekker–CUT&amp;Tag multiome protocol combining direct spatial barcoding of a tissue section with single-cell multiome analysis. This approach enables spatially resolved single-cell profiling of gene expression and the histone H3K27ac modification simultaneously, offering mechanistic insights into gene regulation in the context of tissue microanatomy.

Spatially Resolved, Integrated Single-Cell Multiomic Profiling of the Transcriptome and Epigenomic Targets in Frozen Tissue Sections

Learn reusable single-cell technology for iterative epigenomic analyses using the same cell multiple times. This method enables analysis of multiple epigenetic marks and transcription factors with statistical validation.

Reusable Single Cell for Iterative Epigenomic Analyses

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