Massively parallel single-molecule telomere length measurement with digital real-time PCR

Luo, Yongqiang; Viswanathan, Ramya; Hande, M. Prakash; Loh, Amos Hong Pheng; Cheow, Lih Feng

doi:10.1126/sciadv.abb7944

Cited by 31 publications

(28 citation statements)

References 53 publications

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“…For instance, the metaphase Q-FISH method requires proliferating cells, and U-STELA is not efficient for determining telomeres greater than 8 kb. New approaches for TL analysis based on digital PCR [23] and long-read sequencing [24] have been proposed recently.…”

Section: Telomere Biologymentioning

confidence: 99%

Genetic, Environmental and Lifestyle Determinants of Accelerated Telomere Attrition as Contributors to Risk and Severity of Multiple Sclerosis

et al. 2021

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Telomeres are protective structures at the ends of linear chromosomes. Shortened telomere lengths (TL) are an indicator of premature biological aging and have been associated with a wide spectrum of disorders, including multiple sclerosis (MS). MS is a chronic inflammatory, demyelinating and neurodegenerative disease of the central nervous system. The exact cause of MS is still unclear. Here, we provide an overview of genetic, environmental and lifestyle factors that have been described to influence TL and to contribute to susceptibility to MS and possibly disease severity. We show that several early-life factors are linked to both reduced TL and higher risk of MS, e.g., adolescent obesity, lack of physical activity, smoking and vitamin D deficiency. This suggests that the mechanisms underlying the disease are connected to cellular aging and senescence promoted by increased inflammation and oxidative stress. Additional prospective research is needed to clearly define the extent to which lifestyle changes can slow down disease progression and prevent accelerated telomere loss in individual patients. It is also important to further elucidate the interactions between shared determinants of TL and MS. In future, cell type-specific studies and advanced TL measurement methods could help to better understand how telomeres may be causally involved in disease processes and to uncover novel opportunities for improved biomarkers and therapeutic interventions in MS.

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Section: Telomere Biologymentioning

confidence: 99%

Genetic, Environmental and Lifestyle Determinants of Accelerated Telomere Attrition as Contributors to Risk and Severity of Multiple Sclerosis

et al. 2021

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“…Using a terminal restriction fragment assay (TRF) in Arabidopsis , Hu et al (2015) concluded that RTEL1 loss does not result in telomere shortening whilst Recker et al (2014) showed that the telomeres might even be longer in the mutants. This apparent discrepancy could be addressed using recent high-resolution methods for telomere length measurement ( Lai et al, 2018 ; Kahl et al, 2020 ; Luo et al, 2020 ). More recently, RTEL1 was found to compensate telomere shortening in somatic cells in the absence of telomerase or replication protein A in Arabidopsis mutants ( Olivier et al, 2018 ; Aklilu et al, 2020 ).…”

Section: Discussionmentioning

confidence: 99%

Downregulation of Barley Regulator of Telomere Elongation Helicase 1 Alters the Distribution of Meiotic Crossovers

et al. 2021

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Programmed meiotic DNA double-strand breaks (DSBs), necessary for proper chromosomal segregation and viable gamete formation, are repaired by homologous recombination (HR) as crossovers (COs) or non-crossovers (NCOs). The mechanisms regulating the number and distribution of COs are still poorly understood. The regulator of telomere elongation helicase 1 (RTEL1) DNA helicase was previously shown to enforce the number of meiotic COs in Caenorhabditis elegans but its function in plants has been studied only in the vegetative phase. Here, we characterised barley RTEL1 gene structure and expression using RNA-seq data previously obtained from vegetative and reproductive organs and tissues. Using RNAi, we downregulated RTEL1 expression specifically in reproductive tissues and analysed its impact on recombination using a barley 50k iSelect SNP Array. Unlike in C. elegans, in a population segregating for RTEL1 downregulated by RNAi, high resolution genome-wide genetic analysis revealed a significant increase of COs at distal chromosomal regions of barley without a change in their total number. Our data reveal the important role of RTEL1 helicase in plant meiosis and control of recombination.

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“…The resolution of this method is said to be 0.2kbp -320 kbp. While this assay is a definite improvement over other PCR-based assays, it only provides the average telomere length across all chromosomes and it is not possible to acquire subtelomere-linked and haplotype-resolved data (Luo et al, 2020).…”

Section: Telomere Length Measurement Techniquesmentioning

confidence: 99%

Single-molecule telomere length characterization by optical mapping in nano-channel array: Perspective and review on telomere length measurement

Uppuluri

Varapula

Young

et al. 2021

Environmental Toxicology and Pharmacology

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In humans, the telomere consists of tandem 5′TTAGGG3′ DNA repeats on both ends of all 46 chromosomes. Telomere shortening has been linked to aging and age-related diseases. Similarly, telomere length changes have been associated with chemical exposure, molecular-level DNA damage, and tumor development. Telomere elongation has been associated to tumor development, caused due to chemical exposure and molecular-level DNA damage. The methods used to study these effects mostly rely on average telomere length as a biomarker. The mechanisms regulating subtelomere-specific and haplotype-specific telomere lengths in humans remain understudied and poorly understood, primarily because of technical limitations in obtaining these data for all chromosomes. Recent studies have shown that it is the short telomeres that are crucial in preserving chromosome stability. The identity and frequency of specific critically short telomeres potentially is a useful biomarker for studying aging, age-related diseases, and cancer. Here, we will briefly review the role of telomere length, its measurement, and our recent single-molecule telomere length measurement assay. With this assay, one can measure individual telomere lengths as well as identify their physically linked subtelomeric DNA. This assay can also positively detect telomere loss, characterize novel subtelomeric variants, haploiypes, and previously uncharacterized recombined subtelomeres. We will also discuss its applications in aging cells and cancer cells, highlighting the utility of the single molecule telomere length assay.

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Massively parallel single-molecule telomere length measurement with digital real-time PCR

Cited by 31 publications

References 53 publications

Genetic, Environmental and Lifestyle Determinants of Accelerated Telomere Attrition as Contributors to Risk and Severity of Multiple Sclerosis

Genetic, Environmental and Lifestyle Determinants of Accelerated Telomere Attrition as Contributors to Risk and Severity of Multiple Sclerosis

Downregulation of Barley Regulator of Telomere Elongation Helicase 1 Alters the Distribution of Meiotic Crossovers

Single-molecule telomere length characterization by optical mapping in nano-channel array: Perspective and review on telomere length measurement

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