Visual and fluorometric determination of telomerase activity by using a cationic conjugated polymer and fluorescence resonance energy transfer

Visual and fluorometric determination of telomerase activity by using a cationic conjugated... The detection of telomerase activity is important in cancer diagnosis and in screening for anti-cancer drugs. This work describes a fluorometric method for the determination of telomerase activity by using a cationic conjugated polymer (CCP). Telomerase substrate primers were labelled with carboxyfluorescein (FAM-TS) which display weak electrostatic interactions with the CCP. Hence, fluorescence resonance energy transfer (FRET) from photoexcited CCP to FAM is weak. However, in the presence of telomerase, telomeric repeats (TTAGGG)x were elongated to the 3′-end of FAM-TS to form multiple G-quadruplexes in the presence of potassium ion (K+). These G-quadruplexes trigger strong electrical interaction between the condensed G-quadruplexes and the CCP, and this results in closer proximity and in more efficient FRET. As a result, strong green fluorescence (peaking at 527 nm) is emitted by FAM. Fluorescence can be visually observed under a UV lamp and be used to quantify the activity of telomerase by using a fluorometer. The assay was applied to the determination of HeLa cells in the 30 to 1000 cells per mL range, with a detection limit as low as 5 cells per mL (at an S/N ratio of 3). The method was applied to the detection of various cancer cell lines in human urine samples. The method is simple, sensitive, selective and accurate. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Microchimica Acta Springer Journals

Visual and fluorometric determination of telomerase activity by using a cationic conjugated polymer and fluorescence resonance energy transfer

Loading next page...
 
/lp/springer_journal/visual-and-fluorometric-determination-of-telomerase-activity-by-using-oJ6o0q08pV
Publisher
Springer Vienna
Copyright
Copyright © 2017 by Springer-Verlag GmbH Austria
Subject
Chemistry; Nanochemistry; Nanotechnology; Characterization and Evaluation of Materials; Analytical Chemistry; Microengineering
ISSN
0026-3672
eISSN
1436-5073
D.O.I.
10.1007/s00604-017-2362-5
Publisher site
See Article on Publisher Site

Abstract

The detection of telomerase activity is important in cancer diagnosis and in screening for anti-cancer drugs. This work describes a fluorometric method for the determination of telomerase activity by using a cationic conjugated polymer (CCP). Telomerase substrate primers were labelled with carboxyfluorescein (FAM-TS) which display weak electrostatic interactions with the CCP. Hence, fluorescence resonance energy transfer (FRET) from photoexcited CCP to FAM is weak. However, in the presence of telomerase, telomeric repeats (TTAGGG)x were elongated to the 3′-end of FAM-TS to form multiple G-quadruplexes in the presence of potassium ion (K+). These G-quadruplexes trigger strong electrical interaction between the condensed G-quadruplexes and the CCP, and this results in closer proximity and in more efficient FRET. As a result, strong green fluorescence (peaking at 527 nm) is emitted by FAM. Fluorescence can be visually observed under a UV lamp and be used to quantify the activity of telomerase by using a fluorometer. The assay was applied to the determination of HeLa cells in the 30 to 1000 cells per mL range, with a detection limit as low as 5 cells per mL (at an S/N ratio of 3). The method was applied to the detection of various cancer cell lines in human urine samples. The method is simple, sensitive, selective and accurate.

Journal

Microchimica ActaSpringer Journals

Published: Jun 21, 2017

References

You’re reading a free preview. Subscribe to read the entire article.


DeepDyve is your
personal research library

It’s your single place to instantly
discover and read the research
that matters to you.

Enjoy affordable access to
over 18 million articles from more than
15,000 peer-reviewed journals.

All for just $49/month

Explore the DeepDyve Library

Search

Query the DeepDyve database, plus search all of PubMed and Google Scholar seamlessly

Organize

Save any article or search result from DeepDyve, PubMed, and Google Scholar... all in one place.

Access

Get unlimited, online access to over 18 million full-text articles from more than 15,000 scientific journals.

Your journals are on DeepDyve

Read from thousands of the leading scholarly journals from SpringerNature, Elsevier, Wiley-Blackwell, Oxford University Press and more.

All the latest content is available, no embargo periods.

See the journals in your area

DeepDyve

Freelancer

DeepDyve

Pro

Price

FREE

$49/month
$360/year

Save searches from
Google Scholar,
PubMed

Create lists to
organize your research

Export lists, citations

Read DeepDyve articles

Abstract access only

Unlimited access to over
18 million full-text articles

Print

20 pages / month

PDF Discount

20% off