Toolkit/cell-free chromatin immunoprecipitation

cell-free chromatin immunoprecipitation

Assay Method·Research·Since 2021

Also known as: cfChIP

Taxonomy: Technique Branch / Method. Workflows sit above the mechanism and technique branches rather than replacing them.

Summary

Cell-free chromatin immunoprecipitation (cfChIP) is an assay method that immunoprecipitates histone mark-associated cell-free chromatin from blood plasma. In the cited study, cfChIP targeting H3K36me3-associated cfDNA was used with droplet digital PCR to infer transcriptional activity of specific genes, including EGFR, in the cells that released the cfDNA.

Usefulness & Problems

Why this is useful

cfChIP is useful for extracting transcription-state information from plasma-derived cell-free DNA rather than measuring only sequence variation. The cited work indicates that H3K36me3-associated cfDNA can report whether a particular gene is being transcribed in the source cells and can distinguish enrichment of mutant versus wild-type EGFR fragments in NSCLC plasma.

Problem solved

This method addresses the problem that conventional cfDNA analyses primarily detect genotype but do not directly indicate whether a specific gene is actively transcribed in the cells shedding the DNA. The reported cfChIP approach uses a transcription-associated histone mark, H3K36me3, to infer gene expression status from circulating chromatin fragments.

Problem links

Need tighter control over gene expression timing or amplitude

Derived

Cell-free chromatin immunoprecipitation (cfChIP) is an assay method that immunoprecipitates histone mark-associated cell-free DNA from blood plasma. In the cited study, H3K36me3-associated cfDNA was used to infer whether transcription of a particular gene, including EGFR, was occurring in the cells that released the cfDNA.

Taxonomy & Function

Implementation Constraints

cofactor dependency: cofactor requirement unknownencoding mode: genetically encodedimplementation constraint: context specific validationoperating role: sensor

The reported implementation uses blood plasma as the input material and immunoprecipitation of H3K36me3-associated cell-free chromatin. Detection of allele-specific enrichment was performed by droplet digital PCR comparing EGFR-L858R and EGFR-WT fragments in NSCLC patient samples.

The supplied evidence is limited to a single 2021 study focused on EGFR in non-small-cell lung cancer plasma samples. No broader performance metrics, sensitivity limits, antibody requirements, or validation across additional histone marks, genes, or disease contexts are provided in the evidence.

Validation

Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication

Supporting Sources

Ranked Claims

Claim 1assay capabilitysupports2021Source 1needs review

cfChIP of H3K36me3-associated cfDNA has the potential to delineate whether transcription of a particular gene is occurring in the cells from which its cfDNA originates.

Claim 2assay capabilitysupports2021Source 1needs review

cfChIP of H3K36me3-associated cfDNA has the potential to delineate whether transcription of a particular gene is occurring in the cells from which its cfDNA originates.

Claim 3assay capabilitysupports2021Source 1needs review

cfChIP of H3K36me3-associated cfDNA has the potential to delineate whether transcription of a particular gene is occurring in the cells from which its cfDNA originates.

Claim 4assay capabilitysupports2021Source 1needs review

cfChIP of H3K36me3-associated cfDNA has the potential to delineate whether transcription of a particular gene is occurring in the cells from which its cfDNA originates.

Claim 5assay capabilitysupports2021Source 1needs review

cfChIP of H3K36me3-associated cfDNA has the potential to delineate whether transcription of a particular gene is occurring in the cells from which its cfDNA originates.

Claim 6assay capabilitysupports2021Source 1needs review

cfChIP of H3K36me3-associated cfDNA has the potential to delineate whether transcription of a particular gene is occurring in the cells from which its cfDNA originates.

Claim 7assay capabilitysupports2021Source 1needs review

cfChIP of H3K36me3-associated cfDNA has the potential to delineate whether transcription of a particular gene is occurring in the cells from which its cfDNA originates.

Claim 8assay capabilitysupports2021Source 1needs review

cfChIP of H3K36me3-associated cfDNA has the potential to delineate whether transcription of a particular gene is occurring in the cells from which its cfDNA originates.

Claim 9assay capabilitysupports2021Source 1needs review

cfChIP of H3K36me3-associated cfDNA has the potential to delineate whether transcription of a particular gene is occurring in the cells from which its cfDNA originates.

Claim 10assay capabilitysupports2021Source 1needs review

cfChIP of H3K36me3-associated cfDNA has the potential to delineate whether transcription of a particular gene is occurring in the cells from which its cfDNA originates.

Claim 11assay capabilitysupports2021Source 1needs review

cfChIP of H3K36me3-associated cfDNA has the potential to delineate whether transcription of a particular gene is occurring in the cells from which its cfDNA originates.

Claim 12assay capabilitysupports2021Source 1needs review

cfChIP of H3K36me3-associated cfDNA has the potential to delineate whether transcription of a particular gene is occurring in the cells from which its cfDNA originates.

Claim 13assay capabilitysupports2021Source 1needs review

cfChIP of H3K36me3-associated cfDNA has the potential to delineate whether transcription of a particular gene is occurring in the cells from which its cfDNA originates.

Claim 14assay capabilitysupports2021Source 1needs review

cfChIP of H3K36me3-associated cfDNA has the potential to delineate whether transcription of a particular gene is occurring in the cells from which its cfDNA originates.

Claim 15assay capabilitysupports2021Source 1needs review

cfChIP of H3K36me3-associated cfDNA has the potential to delineate whether transcription of a particular gene is occurring in the cells from which its cfDNA originates.

Claim 16assay capabilitysupports2021Source 1needs review

cfChIP of H3K36me3-associated cfDNA has the potential to delineate whether transcription of a particular gene is occurring in the cells from which its cfDNA originates.

Claim 17assay capabilitysupports2021Source 1needs review

cfChIP of H3K36me3-associated cfDNA has the potential to delineate whether transcription of a particular gene is occurring in the cells from which its cfDNA originates.

Claim 18assay capabilitysupports2021Source 1needs review

cfChIP of H3K36me3-associated cfDNA has the potential to delineate whether transcription of a particular gene is occurring in the cells from which its cfDNA originates.

Claim 19assay capabilitysupports2021Source 1needs review

cfChIP of H3K36me3-associated cfDNA has the potential to delineate whether transcription of a particular gene is occurring in the cells from which its cfDNA originates.

Claim 20assay capabilitysupports2021Source 1needs review

cfChIP of H3K36me3-associated cfDNA has the potential to delineate whether transcription of a particular gene is occurring in the cells from which its cfDNA originates.

Claim 21assay capabilitysupports2021Source 1needs review

cfChIP of H3K36me3-associated cfDNA has the potential to delineate whether transcription of a particular gene is occurring in the cells from which its cfDNA originates.

Claim 22assay capabilitysupports2021Source 1needs review

cfChIP of H3K36me3-associated cfDNA has the potential to delineate whether transcription of a particular gene is occurring in the cells from which its cfDNA originates.

Claim 23assay capabilitysupports2021Source 1needs review

cfChIP of H3K36me3-associated cfDNA has the potential to delineate whether transcription of a particular gene is occurring in the cells from which its cfDNA originates.

Claim 24assay capabilitysupports2021Source 1needs review

cfChIP of H3K36me3-associated cfDNA has the potential to delineate whether transcription of a particular gene is occurring in the cells from which its cfDNA originates.

Claim 25assay capabilitysupports2021Source 1needs review

cfChIP of H3K36me3-associated cfDNA has the potential to delineate whether transcription of a particular gene is occurring in the cells from which its cfDNA originates.

Claim 26assay capabilitysupports2021Source 1needs review

cfChIP of H3K36me3-associated cfDNA has the potential to delineate whether transcription of a particular gene is occurring in the cells from which its cfDNA originates.

Claim 27assay capabilitysupports2021Source 1needs review

cfChIP of H3K36me3-associated cfDNA has the potential to delineate whether transcription of a particular gene is occurring in the cells from which its cfDNA originates.

Claim 28enrichment resultsupports2021Source 1needs review

In blood plasma from NSCLC patients harboring EGFR-L858R, H3K36me3 cfChIP followed by ddPCR revealed significantly higher enrichment of EGFR-L858R fragments than EGFR-WT fragments.

Claim 29enrichment resultsupports2021Source 1needs review

In blood plasma from NSCLC patients harboring EGFR-L858R, H3K36me3 cfChIP followed by ddPCR revealed significantly higher enrichment of EGFR-L858R fragments than EGFR-WT fragments.

Claim 30enrichment resultsupports2021Source 1needs review

In blood plasma from NSCLC patients harboring EGFR-L858R, H3K36me3 cfChIP followed by ddPCR revealed significantly higher enrichment of EGFR-L858R fragments than EGFR-WT fragments.

Claim 31enrichment resultsupports2021Source 1needs review

In blood plasma from NSCLC patients harboring EGFR-L858R, H3K36me3 cfChIP followed by ddPCR revealed significantly higher enrichment of EGFR-L858R fragments than EGFR-WT fragments.

Claim 32enrichment resultsupports2021Source 1needs review

In blood plasma from NSCLC patients harboring EGFR-L858R, H3K36me3 cfChIP followed by ddPCR revealed significantly higher enrichment of EGFR-L858R fragments than EGFR-WT fragments.

Claim 33enrichment resultsupports2021Source 1needs review

In blood plasma from NSCLC patients harboring EGFR-L858R, H3K36me3 cfChIP followed by ddPCR revealed significantly higher enrichment of EGFR-L858R fragments than EGFR-WT fragments.

Claim 34enrichment resultsupports2021Source 1needs review

In blood plasma from NSCLC patients harboring EGFR-L858R, H3K36me3 cfChIP followed by ddPCR revealed significantly higher enrichment of EGFR-L858R fragments than EGFR-WT fragments.

Claim 35enrichment resultsupports2021Source 1needs review

In blood plasma from NSCLC patients harboring EGFR-L858R, H3K36me3 cfChIP followed by ddPCR revealed significantly higher enrichment of EGFR-L858R fragments than EGFR-WT fragments.

Claim 36enrichment resultsupports2021Source 1needs review

In blood plasma from NSCLC patients harboring EGFR-L858R, H3K36me3 cfChIP followed by ddPCR revealed significantly higher enrichment of EGFR-L858R fragments than EGFR-WT fragments.

Claim 37enrichment resultsupports2021Source 1needs review

In blood plasma from NSCLC patients harboring EGFR-L858R, H3K36me3 cfChIP followed by ddPCR revealed significantly higher enrichment of EGFR-L858R fragments than EGFR-WT fragments.

Claim 38enrichment resultsupports2021Source 1needs review

In blood plasma from NSCLC patients harboring EGFR-L858R, H3K36me3 cfChIP followed by ddPCR revealed significantly higher enrichment of EGFR-L858R fragments than EGFR-WT fragments.

Claim 39enrichment resultsupports2021Source 1needs review

In blood plasma from NSCLC patients harboring EGFR-L858R, H3K36me3 cfChIP followed by ddPCR revealed significantly higher enrichment of EGFR-L858R fragments than EGFR-WT fragments.

Claim 40enrichment resultsupports2021Source 1needs review

In blood plasma from NSCLC patients harboring EGFR-L858R, H3K36me3 cfChIP followed by ddPCR revealed significantly higher enrichment of EGFR-L858R fragments than EGFR-WT fragments.

Claim 41enrichment resultsupports2021Source 1needs review

In blood plasma from NSCLC patients harboring EGFR-L858R, H3K36me3 cfChIP followed by ddPCR revealed significantly higher enrichment of EGFR-L858R fragments than EGFR-WT fragments.

Claim 42enrichment resultsupports2021Source 1needs review

In blood plasma from NSCLC patients harboring EGFR-L858R, H3K36me3 cfChIP followed by ddPCR revealed significantly higher enrichment of EGFR-L858R fragments than EGFR-WT fragments.

Claim 43enrichment resultsupports2021Source 1needs review

In blood plasma from NSCLC patients harboring EGFR-L858R, H3K36me3 cfChIP followed by ddPCR revealed significantly higher enrichment of EGFR-L858R fragments than EGFR-WT fragments.

Claim 44enrichment resultsupports2021Source 1needs review

In blood plasma from NSCLC patients harboring EGFR-L858R, H3K36me3 cfChIP followed by ddPCR revealed significantly higher enrichment of EGFR-L858R fragments than EGFR-WT fragments.

Claim 45enrichment resultsupports2021Source 1needs review

In blood plasma from NSCLC patients harboring EGFR-L858R, H3K36me3 cfChIP followed by ddPCR revealed significantly higher enrichment of EGFR-L858R fragments than EGFR-WT fragments.

Claim 46enrichment resultsupports2021Source 1needs review

In blood plasma from NSCLC patients harboring EGFR-L858R, H3K36me3 cfChIP followed by ddPCR revealed significantly higher enrichment of EGFR-L858R fragments than EGFR-WT fragments.

Claim 47enrichment resultsupports2021Source 1needs review

In blood plasma from NSCLC patients harboring EGFR-L858R, H3K36me3 cfChIP followed by ddPCR revealed significantly higher enrichment of EGFR-L858R fragments than EGFR-WT fragments.

Claim 48proof of principlesupports2021Source 1needs review

This study provides proof of principle that cfChIP can be used to identify tumor-specific transcriptional activity of mutated alleles.

Claim 49proof of principlesupports2021Source 1needs review

This study provides proof of principle that cfChIP can be used to identify tumor-specific transcriptional activity of mutated alleles.

Claim 50proof of principlesupports2021Source 1needs review

This study provides proof of principle that cfChIP can be used to identify tumor-specific transcriptional activity of mutated alleles.

Claim 51proof of principlesupports2021Source 1needs review

This study provides proof of principle that cfChIP can be used to identify tumor-specific transcriptional activity of mutated alleles.

Claim 52proof of principlesupports2021Source 1needs review

This study provides proof of principle that cfChIP can be used to identify tumor-specific transcriptional activity of mutated alleles.

Claim 53proof of principlesupports2021Source 1needs review

This study provides proof of principle that cfChIP can be used to identify tumor-specific transcriptional activity of mutated alleles.

Claim 54proof of principlesupports2021Source 1needs review

This study provides proof of principle that cfChIP can be used to identify tumor-specific transcriptional activity of mutated alleles.

Claim 55proof of principlesupports2021Source 1needs review

This study provides proof of principle that cfChIP can be used to identify tumor-specific transcriptional activity of mutated alleles.

Claim 56proof of principlesupports2021Source 1needs review

This study provides proof of principle that cfChIP can be used to identify tumor-specific transcriptional activity of mutated alleles.

Claim 57proof of principlesupports2021Source 1needs review

This study provides proof of principle that cfChIP can be used to identify tumor-specific transcriptional activity of mutated alleles.

Claim 58proof of principlesupports2021Source 1needs review

This study provides proof of principle that cfChIP can be used to identify tumor-specific transcriptional activity of mutated alleles.

Claim 59proof of principlesupports2021Source 1needs review

This study provides proof of principle that cfChIP can be used to identify tumor-specific transcriptional activity of mutated alleles.

Claim 60proof of principlesupports2021Source 1needs review

This study provides proof of principle that cfChIP can be used to identify tumor-specific transcriptional activity of mutated alleles.

Claim 61proof of principlesupports2021Source 1needs review

This study provides proof of principle that cfChIP can be used to identify tumor-specific transcriptional activity of mutated alleles.

Claim 62proof of principlesupports2021Source 1needs review

This study provides proof of principle that cfChIP can be used to identify tumor-specific transcriptional activity of mutated alleles.

Claim 63proof of principlesupports2021Source 1needs review

This study provides proof of principle that cfChIP can be used to identify tumor-specific transcriptional activity of mutated alleles.

Claim 64proof of principlesupports2021Source 1needs review

This study provides proof of principle that cfChIP can be used to identify tumor-specific transcriptional activity of mutated alleles.

Claim 65proof of principlesupports2021Source 1needs review

This study provides proof of principle that cfChIP can be used to identify tumor-specific transcriptional activity of mutated alleles.

Claim 66proof of principlesupports2021Source 1needs review

This study provides proof of principle that cfChIP can be used to identify tumor-specific transcriptional activity of mutated alleles.

Claim 67proof of principlesupports2021Source 1needs review

This study provides proof of principle that cfChIP can be used to identify tumor-specific transcriptional activity of mutated alleles.

Claim 68proof of principlesupports2021Source 1needs review

This study provides proof of principle that cfChIP can be used to identify tumor-specific transcriptional activity of mutated alleles.

Claim 69proof of principlesupports2021Source 1needs review

This study provides proof of principle that cfChIP can be used to identify tumor-specific transcriptional activity of mutated alleles.

Claim 70proof of principlesupports2021Source 1needs review

This study provides proof of principle that cfChIP can be used to identify tumor-specific transcriptional activity of mutated alleles.

Claim 71proof of principlesupports2021Source 1needs review

This study provides proof of principle that cfChIP can be used to identify tumor-specific transcriptional activity of mutated alleles.

Claim 72proof of principlesupports2021Source 1needs review

This study provides proof of principle that cfChIP can be used to identify tumor-specific transcriptional activity of mutated alleles.

Claim 73proof of principlesupports2021Source 1needs review

This study provides proof of principle that cfChIP can be used to identify tumor-specific transcriptional activity of mutated alleles.

Claim 74proof of principlesupports2021Source 1needs review

This study provides proof of principle that cfChIP can be used to identify tumor-specific transcriptional activity of mutated alleles.

Claim 75transcription observationsupports2021Source 1needs review

In representative NSCLC cell lines, both wild-type EGFR and EGFR-L858R are transcribed, and mRNA is similarly expressed per EGFR copy.

Claim 76transcription observationsupports2021Source 1needs review

In representative NSCLC cell lines, both wild-type EGFR and EGFR-L858R are transcribed, and mRNA is similarly expressed per EGFR copy.

Claim 77transcription observationsupports2021Source 1needs review

In representative NSCLC cell lines, both wild-type EGFR and EGFR-L858R are transcribed, and mRNA is similarly expressed per EGFR copy.

Claim 78transcription observationsupports2021Source 1needs review

In representative NSCLC cell lines, both wild-type EGFR and EGFR-L858R are transcribed, and mRNA is similarly expressed per EGFR copy.

Claim 79transcription observationsupports2021Source 1needs review

In representative NSCLC cell lines, both wild-type EGFR and EGFR-L858R are transcribed, and mRNA is similarly expressed per EGFR copy.

Claim 80transcription observationsupports2021Source 1needs review

In representative NSCLC cell lines, both wild-type EGFR and EGFR-L858R are transcribed, and mRNA is similarly expressed per EGFR copy.

Claim 81transcription observationsupports2021Source 1needs review

In representative NSCLC cell lines, both wild-type EGFR and EGFR-L858R are transcribed, and mRNA is similarly expressed per EGFR copy.

Claim 82transcription observationsupports2021Source 1needs review

In representative NSCLC cell lines, both wild-type EGFR and EGFR-L858R are transcribed, and mRNA is similarly expressed per EGFR copy.

Claim 83transcription observationsupports2021Source 1needs review

In representative NSCLC cell lines, both wild-type EGFR and EGFR-L858R are transcribed, and mRNA is similarly expressed per EGFR copy.

Claim 84transcription observationsupports2021Source 1needs review

In representative NSCLC cell lines, both wild-type EGFR and EGFR-L858R are transcribed, and mRNA is similarly expressed per EGFR copy.

Claim 85transcription observationsupports2021Source 1needs review

In representative NSCLC cell lines, both wild-type EGFR and EGFR-L858R are transcribed, and mRNA is similarly expressed per EGFR copy.

Claim 86transcription observationsupports2021Source 1needs review

In representative NSCLC cell lines, both wild-type EGFR and EGFR-L858R are transcribed, and mRNA is similarly expressed per EGFR copy.

Claim 87transcription observationsupports2021Source 1needs review

In representative NSCLC cell lines, both wild-type EGFR and EGFR-L858R are transcribed, and mRNA is similarly expressed per EGFR copy.

Claim 88transcription observationsupports2021Source 1needs review

In representative NSCLC cell lines, both wild-type EGFR and EGFR-L858R are transcribed, and mRNA is similarly expressed per EGFR copy.

Claim 89transcription observationsupports2021Source 1needs review

In representative NSCLC cell lines, both wild-type EGFR and EGFR-L858R are transcribed, and mRNA is similarly expressed per EGFR copy.

Claim 90transcription observationsupports2021Source 1needs review

In representative NSCLC cell lines, both wild-type EGFR and EGFR-L858R are transcribed, and mRNA is similarly expressed per EGFR copy.

Claim 91transcription observationsupports2021Source 1needs review

In representative NSCLC cell lines, both wild-type EGFR and EGFR-L858R are transcribed, and mRNA is similarly expressed per EGFR copy.

Claim 92transcription observationsupports2021Source 1needs review

In representative NSCLC cell lines, both wild-type EGFR and EGFR-L858R are transcribed, and mRNA is similarly expressed per EGFR copy.

Claim 93transcription observationsupports2021Source 1needs review

In representative NSCLC cell lines, both wild-type EGFR and EGFR-L858R are transcribed, and mRNA is similarly expressed per EGFR copy.

Claim 94transcription observationsupports2021Source 1needs review

In representative NSCLC cell lines, both wild-type EGFR and EGFR-L858R are transcribed, and mRNA is similarly expressed per EGFR copy.

Claim 95transcription observationsupports2021Source 1needs review

In representative NSCLC cell lines, both wild-type EGFR and EGFR-L858R are transcribed, and mRNA is similarly expressed per EGFR copy.

Claim 96transcription observationsupports2021Source 1needs review

In representative NSCLC cell lines, both wild-type EGFR and EGFR-L858R are transcribed, and mRNA is similarly expressed per EGFR copy.

Claim 97transcription observationsupports2021Source 1needs review

In representative NSCLC cell lines, both wild-type EGFR and EGFR-L858R are transcribed, and mRNA is similarly expressed per EGFR copy.

Claim 98transcription observationsupports2021Source 1needs review

In representative NSCLC cell lines, both wild-type EGFR and EGFR-L858R are transcribed, and mRNA is similarly expressed per EGFR copy.

Claim 99transcription observationsupports2021Source 1needs review

In representative NSCLC cell lines, both wild-type EGFR and EGFR-L858R are transcribed, and mRNA is similarly expressed per EGFR copy.

Claim 100transcription observationsupports2021Source 1needs review

In representative NSCLC cell lines, both wild-type EGFR and EGFR-L858R are transcribed, and mRNA is similarly expressed per EGFR copy.

Claim 101transcription observationsupports2021Source 1needs review

In representative NSCLC cell lines, both wild-type EGFR and EGFR-L858R are transcribed, and mRNA is similarly expressed per EGFR copy.

Approval Evidence

1 source3 linked approval claimsfirst-pass slug cell-free-chromatin-immunoprecipitation
cell-free chromatin immunoprecipitation (cfChIP)

Source:

assay capabilitysupports

cfChIP of H3K36me3-associated cfDNA has the potential to delineate whether transcription of a particular gene is occurring in the cells from which its cfDNA originates.

Source:

proof of principlesupports

This study provides proof of principle that cfChIP can be used to identify tumor-specific transcriptional activity of mutated alleles.

Source:

transcription observationsupports

In representative NSCLC cell lines, both wild-type EGFR and EGFR-L858R are transcribed, and mRNA is similarly expressed per EGFR copy.

Source:

Comparisons

Source-backed strengths

The cited study reports that H3K36me3 cfChIP has the potential to delineate whether transcription of a particular gene is occurring in the cells from which its cfDNA originates. In blood plasma from NSCLC patients harboring EGFR-L858R, H3K36me3 cfChIP followed by ddPCR showed significantly higher enrichment of EGFR-L858R fragments than EGFR-WT fragments.

cell-free chromatin immunoprecipitation and Affymetrix ATH1 microarray address a similar problem space because they share transcription.

Shared frame: same top-level item type; shared target processes: transcription

Strengths here: looks easier to implement in practice.

Compared with qRT-PCR

cell-free chromatin immunoprecipitation and qRT-PCR address a similar problem space because they share transcription.

Shared frame: same top-level item type; shared target processes: transcription

Strengths here: looks easier to implement in practice.

Compared with RNA sequencing

cell-free chromatin immunoprecipitation and RNA sequencing address a similar problem space because they share transcription.

Shared frame: same top-level item type; shared target processes: transcription

Ranked Citations

  1. 1.
    StructuralSource 1Molecular Oncology2021Claim 1Claim 26Claim 25

    Seeded from load plan for claim c4. Extracted from this source document.