# Standardizing Q-PCR and developing and testing a new DNA-FISH method for high-throughput assessment of telomere length using genomic DNA

> **NIH NIH U01** · GEORGETOWN UNIVERSITY · 2020 · $311,000

## Abstract

ABSTRACT
 With a growing interest in telomere biology in biomedical and epidemiological research, measurement of
telomere length with high precision and accuracy is of paramount importance. Currently, telomere length is
measured in many different laboratories using different methods with no standardization. The different
approaches to measure and report telomere length data hinder comparisons and pooling of data across studies.
Standardization of the most commonly used telomere length measurement method for the field is urgently
needed. Quantitative PCR (Q-PCR) is by far the method most commonly adopted by the epidemiological and
population studies for the measurement of average telomere length. Therefore, we propose to validate and
standardize key components, including pre-PCR and PCR-related factors, of the Q-PCR procedure for a more
reproducible telomere length measurement.
 Despite the low cost and high-throughput nature of the Q-PCR telomere length measurement method,
this approach (and other extant methods) only estimates average telomere length. Average telomere length is
not fully informative of telomere length constitution in cells, because a diploid human cell contains 92
chromosomal ends and telomere lengths at the 92 chromosomal ends are highly heterogeneous. A high
resolution and sensitive method that can measure the length of single telomeres could advance the field by
providing a more detailed assessment of multiple aspects of telomere constitution, not just average telomere
length. In aim 3, we propose to develop a high-throughput DNA fluorescent in situ hybridization (DNA-FISH)
method for the measurement of absolute lengths of single telomeres using genomic DNA. The proposed new
method will generate 4 telomere parameters: 1) average telomere length; 2) telomere length variation, defined
as co-efficient of variation of mean telomere length among all measured telomeres; 3) frequency of short
telomeres; and 4) frequency of long telomeres. This method will provide a new technology to study how
environmental exposures and psychosocial stress impact the changes of not only average telomere length but
also telomere length variation and frequency of short telomeres in the context of aging and disease susceptibility.
 This application is in response to RFA-AG-19-023 “Telomeres as sentinels of environmental exposure,
psychosocial stress, and disease susceptibility: a methods comparison study (U01). Accordingly, we propose
the following three aims: (1) to conduct a collaborative methods comparison study; (2) to collaborate on
developing best practice guidelines for telomere length research; (3) to develop and validate a high-throughput
method to assess telomere length constitution using genomic DNA.

## Key facts

- **NIH application ID:** 10016309
- **Project number:** 5U01ES031786-02
- **Recipient organization:** GEORGETOWN UNIVERSITY
- **Principal Investigator:** YUN-LING ZHENG
- **Activity code:** U01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $311,000
- **Award type:** 5
- **Project period:** 2019-09-12 → 2022-05-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10016309

## Citation

> US National Institutes of Health, RePORTER application 10016309, Standardizing Q-PCR and developing and testing a new DNA-FISH method for high-throughput assessment of telomere length using genomic DNA (5U01ES031786-02). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10016309. Licensed CC0.

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