Real-Time Quantitative PCR of Telomere Length

Gil, Marcel Eduardo; Coetzer, Thérèsa L.

doi:10.1385/mb:27:2:169

Cited by 126 publications

(102 citation statements)

References 9 publications

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“…The telomere length of freshly isolated CD34ϩ cells was measured by real-time quantitative polymerase chain reaction (PCR) analysis according to the method of Cawthon (28), with modifications (29). Genomic DNA was purified using a Wizard Genomic DNA purification kit (Promega, Madison, WI) and quantified according to the copy number of a single-copy gene (36B4).…”

Section: Methodsmentioning

confidence: 99%

Defective proliferative capacity and accelerated telomeric loss of hematopoietic progenitor cells in rheumatoid arthritis

Colmegna

Díaz-Borjón

Fujii

et al. 2008

Arthritis & Rheumatism

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Objective. In rheumatoid arthritis (RA), telomeres of lymphoid and myeloid cells are ageinappropriately shortened, suggesting excessive turnover of hematopoietic precursor cells (HPCs). The purpose of this study was to examine the functional competence (proliferative capacity, maintenance of telomeric reserve) of CD34؉ HPCs in RA.Methods. Frequencies of peripheral blood CD34؉,CD45؉ HPCs from 63 rheumatoid factorpositive RA patients and 48 controls matched for age, sex, and ethnicity were measured by flow cytometry. Proliferative burst, cell cycle dynamics, and induction of lineage-restricted receptors were tested in purified CD34؉ HPCs after stimulation with early hematopoietins. Telomere sequences were quantified by real-time polymerase chain reaction. HPC functions were correlated with the duration, activity, and severity of RA as well as its treatment.Results. In healthy donors, CD34؉ HPCs accounted for 0.05% of nucleated cells; their numbers were strictly age dependent and declined at a rate of 1.3% per year. In RA patients, CD34؉ HPC frequencies were age-independently reduced to 0.03%. Upon growth factor stimulation, control HPCs passed through 5 replication cycles over 4 days. In contrast, RA-derived HPCs completed only 3 generations. Telomeres of RA CD34؉ HPCs were age-inappropriately shortened by 1,600 bp.

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Section: Methodsmentioning

confidence: 99%

Defective proliferative capacity and accelerated telomeric loss of hematopoietic progenitor cells in rheumatoid arthritis

Colmegna

Díaz-Borjón

Fujii

et al. 2008

Arthritis & Rheumatism

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“…11 This method expresses telomere length as a T/S ratio with RNase P as a reference (ABI) for each sample (Supplementary Methods).…”

Section: Telomere Length Measurementmentioning

confidence: 99%

Common variants near TERC are associated with leukocyte telomere length in the Chinese Han population

Shen

Zhou

et al. 2011

Eur J Hum Genet

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A recent genome-wide association study has identified an association between leukocyte telomere length (LTL) and a locus at 3q26 that includes TERC. In order to evaluate the effects of the SNPs rs12696304 and rs16847897 near TERC in the population of mainland China, we conducted an association study of LTL focusing on these two candidate SNPs in a sample of 4016 Chinese Han individuals. Multiple linear regression analyses were performed to evaluate the association of LTL with each SNP adjusted for age, gender and diabetes status. In the study, we confirmed the association of SNP rs12696304 and rs16847897 near TERC with LTL in the Chinese Han population (PB4.5Â10 À3 and 9.5Â10 À5 , respectively). Each copy of the major allele of rs12696304 and rs16847897 was associated with a shorter mean telomere length of 0.024 and 0.031 T/S respectively, which is equivalent to about 3 and 4 years of average age-related telomere attrition. Our short report confirmed the effects of SNPs near TERC on LTL in the Chinese Han population for the first time. Keywords: Chinese population; TERC; leukocyte telomere length INTRODUCTIONTelomeres are structures at the ends of eukaryotic chromosomes, which are made up of a repetitive sequence and which has a major role in genomic stability. Telomere dysregulation can lead to cell death, cell senescence, or abnormal cell proliferation. 1 In humans, leukocyte telomere length (LTL) is getting shorter progressively with age, and is frequently reported to be relatively shorter in aging-related diseases: such as Alzheimer's disease 2 and vascular dementia. 3 Telomere length varies between individuals of the same age, and is found to be inheritable in quantitative-trait linkage analyses of sib pairs. 4,5 Quantitative trait locus studies have mapped putative loci for telomere length to human chromosomes 3p26.1, 10q26.13, 12q12.22 and 14q23.2. [5][6][7] Telomerase is a large, mainly uncharacterized, RNA-protein complex that is responsible for the de novo synthesis and maintenance of telomere ends. TERC is a main component of telomerase, which serves as a template for addition of multiple 6 bp (TTAGGG) telomere repeats. A recent genome-wide association study (GWAS) identified association of common variants near TERC (on 3q26) with telomere length in two large European cohorts of 2917 individuals, followed by replication in three further European cohorts of 9492 individuals. 8 The strongest associations with LTL were observed for SNP rs12696304 and rs16847897 near TERC on 3q26, and the association signals across the 3q26 locus between rs12696304 and rs16847897 are located toward opposite ends of an B87 kb region showing significant

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“…Previously, it was thought that the measurement of telomere by PCR amplification with oligonucleotide primers, TTAGGG and CCCTAA is impossible, because the hybridization may occur between these primers and only primer dimerderived products are generated [74,75]. In their presented study, the primers are modified and are composed of repeated pattern of six bases containing four consecutive paired bases followed by two mismatched bases.…”

Section: Quantitative Polymerase Chain Reaction (Q-pcr)mentioning

confidence: 99%

“…Quantitative-polymerase chain reaction [74,75] Single telomere length analysis [76][77][78] Primer extension/nick translation [3,4] Electron microscopy [79] Telomeric-oligonucleotide ligation assay [80] the measurement of telomere was based on the ratio of telomere to centromere content (TC ratio). This method can measure the minimum of 800 whole cells, or 9 ng of purified DNA, and thus increased the assay sensitivity and DNA isolation is not needed when the cell numbers are too small.…”

Section: Modifications Referencesmentioning

confidence: 99%

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The telomere length dynamic and methods of its assessment

Lin

Yan

2005

J Cellular Mol Med

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Human telomeres are composed of long repeating sequences of TTAGGG, associated with a variety of telomere‐binding proteins. Its function as an end‐protector of chromosomes prevents the chromosome from end‐to‐end fusion, recombination and degradation. Telomerase acts as reverse transcriptase in the elongation of telomeres, which prevent the loss of telomeres due to the end replication problems. However, telomerase activity is detected at low level in somatic cells and high level in embryonic stem cells and tumor cells. It confers immortality to embryonic stem cells and tumor cells. In most tumor cells, telomeres are extremely short and stable. Telomere length is an important indicator of the telomerase activity in tumor cells and it may be used in the prognosis of malignancy. Thus, the assessment of telomeres length is of great experimental and clinical significance. This review describes the role of telomere and telomerase in cancer pathogenesis and the dynamics of the telomeres length in different cell types. The various methods of measurement of telomeres length, i.e. southern blot, hybridization protection assay, fluorescence in situ hybridization, primed in situ, quantitative PCR and single telomere length analysis are discussed. The principle and comparative evaluation of these methods are reviewed. The detection of G‐strand overhang by telomeric‐oligonucleotide ligation assay, primer extension/nick translation assay and electron microscopy are briefly discussed.

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Real-Time Quantitative PCR of Telomere Length

Cited by 126 publications

References 9 publications

Defective proliferative capacity and accelerated telomeric loss of hematopoietic progenitor cells in rheumatoid arthritis

Defective proliferative capacity and accelerated telomeric loss of hematopoietic progenitor cells in rheumatoid arthritis

Common variants near TERC are associated with leukocyte telomere length in the Chinese Han population

The telomere length dynamic and methods of its assessment

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