An Integrated Workflow to Study the Promoter-Centric Spatio-Temporal Genome Architecture in Scarce Cell Populations

Lloren&#231; Rovirosa; Laureano Tom&#225;s-Daza; Blanca Urmeneta; Alfonso Valencia; Biola M. Javierre

doi:10.3791/65316

Journal

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Genetics

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An Integrated Workflow to Study the Promoter-Centric Spatio-Temporal Genome Architecture in Scarce Cell Populations

JoVE Journal
Genetics

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JoVE Journal Genetics

An Integrated Workflow to Study the Promoter-Centric Spatio-Temporal Genome Architecture in Scarce Cell Populations

1,915 Views

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11:36 min

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April 21, 2023

DOI:

10.3791/65316-v

DOI:

10.3791/65316-v

•

11:36 min

April 21, 2023

•

4 Views

Llorenç Rovirosa*¹, Laureano Tomás-Daza*^1,2, Blanca Urmeneta¹, Alfonso Valencia^2,3, Biola M. Javierre^1,4

¹Josep Carreras Leukaemia Research Institute, ²Barcelona Supercomputing Center, ³Catalan Institution for Research and Advanced Studies (ICREA), ⁴Institute for Health Science Research Germans Trias i Pujol

Transcript

With liCHi-C, the main question that we are trying to answer is how the chromatin interacts in the nucleus, and therefore, to understand a new layer of gene regulation. We have proven with liCHi-C that we can explore the genome organization of a given sample, link the non-coding mutation associated with disease with potential genes affected, and detect chromosomal rearrangements, such as translocation, for example, in tumor samples. Now we can explore the 3D chromatin organization of scar cell types, such as primary stem cells or relevant clinical samples.

Our protocol reduces the input materials and the time and cost needed to obtain quality data to explore the genome organization at the restriction fragment resolution. liCHi-C is expanding the reach of the 3D chromatin organization field, allowing us to explore new cell types previously impossible to analyze. Thanks to liCHi-C, the scientific community can explore the special regulation of gene expression.

For example, in malignant transformation only in a specific clonal sub-population inside a tumor.

Summary

Gene expression is regulated by interactions of gene promoters with distal regulatory elements. Here, we descirbe how low input Capture Hi-C (liCHi-C) allows the identification of these interactions in rare cell types, which were previously unmeasurable.