Imaging-Based In Situ Analysis of 5-Methylcytosine at Low Repetitive Single Gene Loci with Transcription-Activator-Like Effector Probes.
Journal
ACS chemical biology
ISSN: 1554-8937
Titre abrégé: ACS Chem Biol
Pays: United States
ID NLM: 101282906
Informations de publication
Date de publication:
17 02 2023
17 02 2023
Historique:
pubmed:
25
1
2023
medline:
22
2
2023
entrez:
24
1
2023
Statut:
ppublish
Résumé
Transcription-activator-like effectors (TALEs) are programmable DNA binding proteins that can be used for sequence-specific, imaging-based analysis of cellular 5-methylcytosine. However, this has so far been limited to highly repetitive satellite DNA. To expand this approach to the analysis of coding single gene loci, we here explore a number of signal amplification strategies for increasing imaging sensitivity with TALEs. We develop a straightforward amplification protocol and employ it to target the MUC4 gene, which features only a small cluster of repeat sequences. This offers high sensitivity imaging of MUC4, and in costaining experiments with pairs of one TALE selective for unmethylated cytosine and one universal control TALE enables analyzing methylation changes in the target independently of changes in target accessibility. These advancements offer prospects for 5-methylcytosine analysis at coding, nonrepetitive gene loci by the use of designed TALE probe collections.
Identifiants
pubmed: 36693632
doi: 10.1021/acschembio.2c00857
pmc: PMC9942090
doi:
Substances chimiques
Transcription Activator-Like Effectors
0
5-Methylcytosine
6R795CQT4H
DNA
9007-49-2
DNA-Binding Proteins
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
230-236Références
Science. 2012 Feb 10;335(6069):716-9
pubmed: 22223736
Chem Commun (Camb). 2016 Dec 6;52(99):14238-14241
pubmed: 27872906
Nature. 2002 Apr 4;416(6880):552-6
pubmed: 11932749
Cell. 2013 Dec 19;155(7):1479-91
pubmed: 24360272
Proc Natl Acad Sci U S A. 2013 Dec 24;110(52):21048-53
pubmed: 24324157
Nat Struct Mol Biol. 2013 Nov;20(11):1321-4
pubmed: 24096363
Nat Rev Genet. 2012 Oct;13(10):679-92
pubmed: 22945394
Nature. 2012 Sep 6;489(7414):57-74
pubmed: 22955616
Cell. 2014 Oct 23;159(3):635-46
pubmed: 25307933
Oncotarget. 2016 Jul 5;7(27):42553-42565
pubmed: 27283771
Nat Commun. 2017 Sep 21;8(1):649
pubmed: 28935858
Nucleic Acids Res. 2013 Oct;41(19):e186
pubmed: 23990328
J Mol Biol. 2013 Feb 8;425(3):479-91
pubmed: 23220192
Nat Commun. 2017 Mar 14;8:14725
pubmed: 28290446
Angew Chem Int Ed Engl. 2020 Jun 2;59(23):8927-8931
pubmed: 32167219
Curr Opin Chem Biol. 2021 Aug;63:1-10
pubmed: 33588304
FASEB J. 2008 Apr;22(4):966-81
pubmed: 18024835
J Am Chem Soc. 2015 Jan 14;137(1):2-5
pubmed: 25562518
Science. 2022 Apr;376(6588):44-53
pubmed: 35357919
Nat Rev Genet. 2016 Aug;17(8):487-500
pubmed: 27346641
Wellcome Open Res. 2021 Oct 13;6:265
pubmed: 34796278
Nucleic Acids Res. 2008 Feb;36(2):423-34
pubmed: 18039709
Genome Biol. 2016 Apr 18;17:72
pubmed: 27091476
ACS Synth Biol. 2018 Jan 19;7(1):176-186
pubmed: 28849913
J Am Chem Soc. 2018 May 9;140(18):5904-5908
pubmed: 29677450
ACS Chem Biol. 2016 Dec 16;11(12):3294-3299
pubmed: 27978710
Nat Commun. 2017 Oct 12;8(1):901
pubmed: 29026078
Nucleic Acids Res. 2014 Apr;42(6):e38
pubmed: 24371265
Cell Res. 2012 Oct;22(10):1502-4
pubmed: 22945353
Cytogenet Genome Res. 2007;118(1):42-54
pubmed: 17901699