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Updated: Sep 4, 2025
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Immunofluorescence Analysis of Endogenous and Exogenous Centromere-kinetochore Proteins
Published on: March 3, 2016
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Enrichment of centromeric DNA from human cells
Riccardo Gamba1, Giulia Mazzucco2, Therese Wilhelm1
1Institut Curie, PSL Research University, CNRS, UMR 144, Paris, France.
Plos Genetics
|July 19, 2022
Summary
Centromeres, crucial for chromosome segregation, are difficult to study due to their repetitive nature. The new CenRICH technique enriches centromeric DNA, making its sequencing more affordable and efficient for genetic and epigenetic analysis.
Area of Science:
- Genomics
- Epigenetics
- Molecular Biology
Background:
- Centromeres are essential for accurate chromosome segregation during cell division.
- Canonical centromeres comprise large tandem repetitive DNA arrays, posing challenges for genetic and epigenetic studies.
- High-coverage sequencing of centromeric regions is costly and technically demanding.
Purpose of the Study:
- To develop a cost-effective and efficient method for enriching centromeric DNA from human cells.
- To facilitate the genetic and epigenetic analysis of centromeric and pericentromeric regions.
- To enable advanced single-molecule studies of centromeric DNA.
Main Methods:
- Developed CenRICH (Centromere Enrichment by Restriction enzyme digestion and size Fractionation).
- Utilized selective restriction digestion and size fractionation (>20 kb) to isolate centromeric DNA.
- Combined CenRICH with long-read sequencing and DNA combing for methylation and fiber analysis.
Main Results:
- Achieved over 25-fold enrichment of centromeric DNA.
- Enriched samples contained up to 60% centromeric repeats.
- Demonstrated successful investigation of DNA methylation and single-molecule replication of centromeric fibers.
Conclusions:
- CenRICH significantly enhances the proportion of centromeric DNA in sequencing samples.
- This method offers a more affordable and efficient approach to studying centromeric sequences and their epigenetic modifications.
- CenRICH is a valuable tool for advancing research on chromosome segregation and genome stability.
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