Rapid method for mean telomere length measurement directly from cell lysates.
Telomere length is involved in cell survival, tumorigenesis, and early aging. We present here an innovative method to determine the mean telomere length without any DNA purification. Our strategy is to measure both the DNA concentration and the number of telomeric units (TTAGGG) directly from cell lysate produced by the combined action of NaOH (pH>13) and sonication directly on cell pellet. Telomere units are quantified using an enzyme hybridization assay on 96-well microtiter plates grafted with a captor sequence. A biotin-coupled-tracer oligonucleotide hybridizes with telomere fragments and the enzymatic reaction is performed with a streptavidin-acetylcholinesterase conjugate, using the colorimetric method of Ellman. OD measure is directly proportional to the number of telomere units in cell lysate. This scalable technique allows the determination of mean telomere length simultaneously in many samples. This assay will be highly efficient to screen new drugs involved in chemotherapy targeting telomerase or directly telomeres.
Freulet-Marriere
M A
MA
CEA-FAR, DSV/DRR/LRO, 18, Route du Panorama-BP6, 92265, Fontenay aux Roses, Cedex, France.
Potocki-Veronese
G
G
Deverre
J R
JR
Sabatier
L
L
D015139:Blotting, Southern; D002460:Cell Line; D006801:Humans; D009693:Nucleic Acid Hybridization; D016615:Telomere