(Po)STAC (Polycistronic SunTAg modified CRISPR) enables live-cell and fixed-cell super-resolution imaging of multiple genes

CRISPR/dCas9-based labeling has allowed direct visualization of genomic regions in living cells. However, poor labeling efficiency and signal-to-background ratio have limited its application to visualize genome organization using super-resolution microscopy. We developed (Po)STAC (Polycistronic SunT...

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Detalles Bibliográficos
Autores principales: Neguembor, Maria V, Sebastian-Perez, Ruben, Aulicino, Francesco, Gomez-Garcia, Pablo A, Cosma, Maria P, Lakadamyali, Melike
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2018
Materias:
Methods Online
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5861460/
https://www.ncbi.nlm.nih.gov/pubmed/29294098
http://dx.doi.org/10.1093/nar/gkx1271
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author Neguembor, Maria V
Sebastian-Perez, Ruben
Aulicino, Francesco
Gomez-Garcia, Pablo A
Cosma, Maria P
Lakadamyali, Melike
author_facet Neguembor, Maria V
Sebastian-Perez, Ruben
Aulicino, Francesco
Gomez-Garcia, Pablo A
Cosma, Maria P
Lakadamyali, Melike
author_sort Neguembor, Maria V
collection PubMed
description CRISPR/dCas9-based labeling has allowed direct visualization of genomic regions in living cells. However, poor labeling efficiency and signal-to-background ratio have limited its application to visualize genome organization using super-resolution microscopy. We developed (Po)STAC (Polycistronic SunTAg modified CRISPR) by combining CRISPR/dCas9 with SunTag labeling and polycistronic vectors. (Po)STAC enhances both labeling efficiency and fluorescence signal detected from labeled loci enabling live cell imaging as well as super-resolution fixed-cell imaging of multiple genes with high spatiotemporal resolution.
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spelling pubmed-58614602018-03-28 (Po)STAC (Polycistronic SunTAg modified CRISPR) enables live-cell and fixed-cell super-resolution imaging of multiple genes Neguembor, Maria V Sebastian-Perez, Ruben Aulicino, Francesco Gomez-Garcia, Pablo A Cosma, Maria P Lakadamyali, Melike Nucleic Acids Res Methods Online CRISPR/dCas9-based labeling has allowed direct visualization of genomic regions in living cells. However, poor labeling efficiency and signal-to-background ratio have limited its application to visualize genome organization using super-resolution microscopy. We developed (Po)STAC (Polycistronic SunTAg modified CRISPR) by combining CRISPR/dCas9 with SunTag labeling and polycistronic vectors. (Po)STAC enhances both labeling efficiency and fluorescence signal detected from labeled loci enabling live cell imaging as well as super-resolution fixed-cell imaging of multiple genes with high spatiotemporal resolution. Oxford University Press 2018-03-16 2017-12-23 /pmc/articles/PMC5861460/ /pubmed/29294098 http://dx.doi.org/10.1093/nar/gkx1271 Text en © The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research. http://creativecommons.org/licenses/by-nc/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com
spellingShingle Methods Online
Neguembor, Maria V
Sebastian-Perez, Ruben
Aulicino, Francesco
Gomez-Garcia, Pablo A
Cosma, Maria P
Lakadamyali, Melike
(Po)STAC (Polycistronic SunTAg modified CRISPR) enables live-cell and fixed-cell super-resolution imaging of multiple genes
title (Po)STAC (Polycistronic SunTAg modified CRISPR) enables live-cell and fixed-cell super-resolution imaging of multiple genes
title_full (Po)STAC (Polycistronic SunTAg modified CRISPR) enables live-cell and fixed-cell super-resolution imaging of multiple genes
title_fullStr (Po)STAC (Polycistronic SunTAg modified CRISPR) enables live-cell and fixed-cell super-resolution imaging of multiple genes
title_full_unstemmed (Po)STAC (Polycistronic SunTAg modified CRISPR) enables live-cell and fixed-cell super-resolution imaging of multiple genes
title_short (Po)STAC (Polycistronic SunTAg modified CRISPR) enables live-cell and fixed-cell super-resolution imaging of multiple genes
title_sort (po)stac (polycistronic suntag modified crispr) enables live-cell and fixed-cell super-resolution imaging of multiple genes
topic Methods Online
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5861460/
https://www.ncbi.nlm.nih.gov/pubmed/29294098
http://dx.doi.org/10.1093/nar/gkx1271
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