Assessing Telomere Length Using Surface Enhanced Raman Scattering

Abstract

Telomere length can provide valuable insight into telomeres and telomerase related diseases, including cancer. Here, we present a brand-new optical telomere length measurement protocol using surface enhanced Raman scattering (SERS). In this protocol, two single strand DNA are used as SERS probes. They are labeled with two different Raman molecules and can specifically hybridize with telomeres and centromere, respectively. First, genome DNA is extracted from cells. Then the telomere and centromere SERS probes are added into the genome DNA. After hybridization with genome DNA, excess SERS probes are removed by magnetic capturing nanoparticles. Finally, the genome DNA with SERS probes attached is dropped onto a SERS substrate and subjected to SERS measurement. Longer telomeres result in more attached telomere probes, thus a stronger SERS signal. Consequently, SERS signal can be used as an indicator of telomere length. Centromere is used as the inner control. By calibrating the SERS intensity of telomere probe with that of the centromere probe, SERS based telomere measurement is realized. This protocol does not require polymerase chain reaction (PCR) or electrophoresis procedures, which greatly simplifies the detection process. We anticipate that this easy-operation and cost-effective protocol is a fine alternative for the assessment of telomere length.

Advances in surface‐enhanced Raman spectroscopy for cancer diagnosis and staging

“…Conventional methods for telomere length detection include southern blot, quantitative PCR, fluorescence in situ hybridization, and hybridization protection assay. All of these strategies suffer from shortcoming of sophisticated protocols, high costs, and relatively low accuracy . On the other hand, SERS‐based detection of telomerase will be advantageous over PCR‐free and enzyme‐free detection of telomerase and real‐time monitoring.…”

Short Telomere Syndromes in Clinical Practice: Bridging Bench and Bedside

“…Several methods have been described to measure telomere length, namely fluorescence in-situ hybridization or FISH [Flow FISH and Quantitative (Q)-FISH 28–30 ), quantitative polymerase chain reaction or Q PCR-based techniques 31 (including single-cell telomere length measurement 32 ), monochrome multiplex Q PCR (MMQ PCR), optical techniques (surface enhanced Raman scattering 33 ), hybridization protection assay 34 and telomere restriction fragment length analysis. 35 Although no single technique is ideal for use in all clinical and research scenarios, the telomere restriction fragment length analysis is considered the ‘gold standard’ largely due to the simplicity of its protocol and accuracy, however its widespread use is limited as it requires large amounts of DNA for analysis.…”

Surface Enhanced Raman Scattering Based in Situ Hybridization Strategy for Telomere Length Assessment

“…Up to now, there have been several kinds of well established telomere length measurement methods, such as Southern blot, , quantitative PCR (qPCR), , hybridization protection assay (HPA), and fluorescent in situ hybridization (FISH). ,, The features of these methods have been summarized in our previous work . Among these methods, FISH is the only one which can analyze the telomere length of a single cell or a single chromosome, while the other methods only provide averaged telomere length of the whole genome.…”

“…, metal nanoparticles) ensures that SERS has a fine sensitivity . Using SERS technique, we have previously developed a SERS based telomere length measurement protocol . However, like most of the other methods, it can only measure averaged telomere length of many cells.…”