Methods for the determination of telomere length in a...

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid

Reexamination Certificate

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C435S287200, C536S024310

Reexamination Certificate

active

08084203

ABSTRACT:
The invention relates to methods and reagents for the determination of telomere length in tissue sections by the single cell telomeric mapping technique based on a fluorescent in situ hybridization step using a telomere-specific probe and an interpolation step using a standard curve correlating fluorescent intensity and telomere length obtained from a collection of cell lines of known telomere length. The invention further relates to methods for the identification of stem cell niches within tissues and for the identification of compounds capable of triggering stem cell mobilization using the telomere length as criteria for the identification of stem cells and which rely on the single cell telomeric mapping technique of the invention.

REFERENCES:
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patent: 2004/035597 (2004-04-01), None
patent: 2008081451 (2008-07-01), None
Canela, A. et al. Proceedings of the National Academy of Sciences USA 104(13):5300-5305 (Mar. 27, 2007; published online Mar. 16, 2007).
Baerlocher, G.M. et al. Nature Protocols 1(5):2365-2376 (published online Dec. 21, 2006).
Andres Canela Rodriguez, Estudio de las nuevas funciones de la telomerasa en la tumorigenesis, Thesis Doctoral 2007, pp. 1-257 (cited in 8 parts), publically available Jan. 2008.
Ignacio Flores, et al., The longest telomeres: a general signature of adult stem cell compartments, GENES & DEVELEPMENT, pp. 654-667, vol. 22, Mar. 1, 2008 (online Feb. 18, 2008).
Steven S.S. Poon, et al.,Telonnere Length Measurements Using Digital Fluorescence Microscopy, Cytometry, 1999 pp. 267-278, vol. 36.
Sabita N. Saldanha, et al., Assesment of telomere length and factors that contribute to its stability, Eur. J. Biochem, 2003, pp. 389-403, vol. 270.
Irene Siegl-Cachedenier, et al., Telomerase reverses epidermal hair follicle stem cell defects and loss of long-term survival associated with critically short telomeres, The Journal of Cell Biology, Oct. 2, 2007, pp. 277-290, vol. 179, No. 2.
Ignacio Flores, et al., Telomerase regulation and stem cell behaviour, Current Opinion in Cell Biology, 2006, pp. 254-260, vol. 18
English translation of Conclusion from Andes Canaela Rodriguez Doctoral Thesis, 2007, (3 pages).
Canela, A., et al., “Constitutive expression of tert in thymocytes leads to increased incidence and dissemination of T-cell lymphoma in Lck-Tert mice,” Mol. Cell Biol. May 2004; 24(10):4275-93.
Canela, A., et al., “High-throughput telomere length quantification by FISH and its application to human population studies,” Proc. Natl. Acad. Sci. USA Mar. 27, 2007; 104(13):5300-5.
Flores, I., “The longest telomeres: a general signature of adult stem cell compartments”. 2007 Sometido, 43 page manuscript (vol./No./pp. N/A).

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