A siRNA-based screen for genes involved in chromosome end protection.

Telomeres are nucleoprotein complexes which protect the ends of linear chromosomes from detection as DNA damage and provide a sequence buffer against replication-associated shortening. In mammals, telomeres consist of repetitive DNA sequence (TTAGGG) and associated proteins. The telomeric core compl...

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Autores principales:	Daniel H Lackner, Daniel Durocher, Jan Karlseder
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2011
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/5db12512146547bcbd525592bbb9bbe5
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spelling	oai:doaj.org-article:5db12512146547bcbd525592bbb9bbe52021-11-18T06:51:22ZA siRNA-based screen for genes involved in chromosome end protection.1932-620310.1371/journal.pone.0021407https://doaj.org/article/5db12512146547bcbd525592bbb9bbe52011-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/21760879/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203Telomeres are nucleoprotein complexes which protect the ends of linear chromosomes from detection as DNA damage and provide a sequence buffer against replication-associated shortening. In mammals, telomeres consist of repetitive DNA sequence (TTAGGG) and associated proteins. The telomeric core complex is called shelterin and is comprised of the proteins TRF1, TRF2, POT1, TIN2, TPP1 and RAP1. Excessive telomere shortening or de-protection of telomeres through the loss of shelterin subunits allows the detection of telomeres as DNA damage, which can be visualized as DNA damage protein foci at chromosome ends called TIF (Telomere Dysfunction-Induced Foci). We sought to exploit the TIF phenotype as marker for telomere dysfunction to identify novel genes involved in telomere protection by siRNA-mediated knock-down of a set of 386 candidates. Here we report the establishment, specificity and feasibility of such a screen and the results of the genes tested. Only one of the candidate genes showed a unique TIF phenotype comparable to the suppression of the main shelterin components TRF2 or TRF1 and that gene was identified as a TRF1-like pseudogene. We also identified a weak TIF phenotype for SKIIP (SNW1), a splicing factor and transcriptional co-activator. However, the knock-down of SKIIP also induced a general, not telomere-specific DNA damage response, which complicates conclusions about a telomeric role. In summary, this report is a technical demonstration of the feasibility of a cell-based screen for telomere deprotection with the potential of scaling it to a high-throughput approach.Daniel H LacknerDaniel DurocherJan KarlsederPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 6, Iss 6, p e21407 (2011)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Daniel H Lackner Daniel Durocher Jan Karlseder A siRNA-based screen for genes involved in chromosome end protection.
description	Telomeres are nucleoprotein complexes which protect the ends of linear chromosomes from detection as DNA damage and provide a sequence buffer against replication-associated shortening. In mammals, telomeres consist of repetitive DNA sequence (TTAGGG) and associated proteins. The telomeric core complex is called shelterin and is comprised of the proteins TRF1, TRF2, POT1, TIN2, TPP1 and RAP1. Excessive telomere shortening or de-protection of telomeres through the loss of shelterin subunits allows the detection of telomeres as DNA damage, which can be visualized as DNA damage protein foci at chromosome ends called TIF (Telomere Dysfunction-Induced Foci). We sought to exploit the TIF phenotype as marker for telomere dysfunction to identify novel genes involved in telomere protection by siRNA-mediated knock-down of a set of 386 candidates. Here we report the establishment, specificity and feasibility of such a screen and the results of the genes tested. Only one of the candidate genes showed a unique TIF phenotype comparable to the suppression of the main shelterin components TRF2 or TRF1 and that gene was identified as a TRF1-like pseudogene. We also identified a weak TIF phenotype for SKIIP (SNW1), a splicing factor and transcriptional co-activator. However, the knock-down of SKIIP also induced a general, not telomere-specific DNA damage response, which complicates conclusions about a telomeric role. In summary, this report is a technical demonstration of the feasibility of a cell-based screen for telomere deprotection with the potential of scaling it to a high-throughput approach.
format	article
author	Daniel H Lackner Daniel Durocher Jan Karlseder
author_facet	Daniel H Lackner Daniel Durocher Jan Karlseder
author_sort	Daniel H Lackner
title	A siRNA-based screen for genes involved in chromosome end protection.
title_short	A siRNA-based screen for genes involved in chromosome end protection.
title_full	A siRNA-based screen for genes involved in chromosome end protection.
title_fullStr	A siRNA-based screen for genes involved in chromosome end protection.
title_full_unstemmed	A siRNA-based screen for genes involved in chromosome end protection.
title_sort	sirna-based screen for genes involved in chromosome end protection.
publisher	Public Library of Science (PLoS)
publishDate	2011
url	https://doaj.org/article/5db12512146547bcbd525592bbb9bbe5
work_keys_str_mv	AT danielhlackner asirnabasedscreenforgenesinvolvedinchromosomeendprotection AT danieldurocher asirnabasedscreenforgenesinvolvedinchromosomeendprotection AT jankarlseder asirnabasedscreenforgenesinvolvedinchromosomeendprotection AT danielhlackner sirnabasedscreenforgenesinvolvedinchromosomeendprotection AT danieldurocher sirnabasedscreenforgenesinvolvedinchromosomeendprotection AT jankarlseder sirnabasedscreenforgenesinvolvedinchromosomeendprotection
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A siRNA-based screen for genes involved in chromosome end protection.

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