# Bridge-seq, a new tool to analyze human genome segregation defects

> **NIH NIH R21** · NEW YORK UNIVERSITY · 2022 · $199,427

## Abstract

PROJECT SUMMARY
Defects in chromosome segregation are detrimental to genome integrity and is a hallmark of cancer. A
common phenotype of chromosome segregation problems is the presence of DAPI stained DNA between
segregating chromosome masses in anaphase. These “anaphase bridges” are poorly characterized, in part
due to the assumption that their content is invariant or random. This assumption may be false because
previous studies have identified mutants that lead to bridges specifically enriched in centromeric, telomeric or
ribosomal DNA. These were candidate sequences analyzed using fluorescence in situ hybridization (FISH) or
immunofluorescence analyses of proteins that bind to them. To date, an unbiased method to determine the
complement and frequency of human genomic sequences contained in the anaphase bridges is not available.
Knowing the primary effect of chromosome segregation defects at the sequence level will enable connecting
them to the large sequencing projects characterizing genome integrity problems in cancers and aging. For
instance, if certain cancer-associated mutations skew towards specific aneuploidies, their consequences may
be cell-type specific based on the genes that are affected.
The critical barrier to determining the DNA content of the bridges is a technical limitation. Here, we propose an
innovative combination of two technologies to develop Bridge-seq: laser capture microdissection (LCM) to
isolate DNA at anaphase bridges and high-throughput sequencing to identify genomic sequences. We will
develop Bridge-seq and test its feasibility in multiple systems affecting different regions of the genome based
on mutations in normal, cancer, and aging cells. In aim 1, We will establish the conditions for Bridge-seq using
conditions that allow for a robust induction of DAPI-staining bridges that are enriched for rDNA. In aim 2, to
assess the validity of using Bridge-seq to study anaphase bridge content across a range of conditions and
different mutations, we will analyze bridges from mutants shown to effect other regions of chromosomes,
including telomeres and centromeres. In Aim 3 we will apply Bridge-seq to analyze and compare anaphase
bridges in aging and cancer cells. Our unbiased analysis will provide new insights into the nature of human
genomic sequences that are vulnerable to segregation defects and that ultimately drive genomic instability.

## Key facts

- **NIH application ID:** 10456922
- **Project number:** 5R21AG070425-02
- **Recipient organization:** NEW YORK UNIVERSITY
- **Principal Investigator:** Sevinc Ercan
- **Activity code:** R21 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $199,427
- **Award type:** 5
- **Project period:** 2021-08-01 → 2024-04-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10456922

## Citation

> US National Institutes of Health, RePORTER application 10456922, Bridge-seq, a new tool to analyze human genome segregation defects (5R21AG070425-02). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10456922. Licensed CC0.

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