Journal Article

DKFZ-2022-00325

http://join2-wiki.gsi.de/foswiki/pub/Main/Artwork/join2_logo100x88.png ALT-FISH quantifies alternative lengthening of telomeres activity by imaging of single-stranded repeats.

Frank, L. (First author)DKFZ* ; Rademacher, A.DKFZ* ; Mücke, N.DKFZ* ; Tirier, S. M.DKFZ* ; Koeleman, E.DKFZ* ; Knotz, C.DKFZ* ; Schumacher, S.DKFZ* ; Stainczyk, S. A.DKFZ* ; Westermann, F.DKFZ* ; Fröhling, S.DKFZ* ; Chudasama, P.DKFZ* ; Rippe, K. (Last author)DKFZ*

2022
Oxford Univ. Press Oxford

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Please use a persistent id in citations: doi:10.1093/nar/gkac113

Abstract: Alternative lengthening of telomeres (ALT) occurs in ∼10% of cancer entities. However, little is known about the heterogeneity of ALT activity since robust ALT detection assays with high-throughput in situ readouts are lacking. Here, we introduce ALT-FISH, a method to quantitate ALT activity in single cells from the accumulation of single-stranded telomeric DNA and RNA. It involves a one-step fluorescent in situ hybridization approach followed by fluorescence microscopy imaging. Our method reliably identified ALT in cancer cell lines from different tumor entities and was validated in three established models of ALT induction and suppression. Furthermore, we successfully applied ALT-FISH to spatially resolve ALT activity in primary tissue sections from leiomyosarcoma and neuroblastoma tumors. Thus, our assay provides insights into the heterogeneity of ALT tumors and is suited for high-throughput applications, which will facilitate screening for ALT-specific drugs.

Note: #EA:B066#LA:B066# / 2022 Jun 24;50(11):e61

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Contributing Institute(s):

1. B066 Chromatin-Netzwerke (B066)
2. B087 Neuroblastom Genomik (B087)
3. Translationale Medizinische Onkologie (B340)
4. DKTK HD zentral (HD01)
5. Präzisions-Sarkomforschung (B390)

Research Program(s):

1. 312 - Funktionelle und strukturelle Genomforschung (POF4-312) (POF4-312)

Appears in the scientific report 2022

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