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

Maria V Neguembor; Ruben Sebastian-Perez; Francesco Aulicino; Pablo A Gomez-Garcia; Maria P Cosma; Melike Lakadamyali

doi:10.1093/nar/gkx1271

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

Maria V Neguembor, Ruben Sebastian-Perez, Francesco Aulicino, Pablo A Gomez-Garcia, Maria P Cosma, Melike Lakadamyali

School of Biochemistry

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Abstract

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.

Original language	English
Article number	e30
Number of pages	10
Journal	Nucleic Acids Research
Volume	46
Issue number	5
DOIs	https://doi.org/10.1093/nar/gkx1271
Publication status	Published - 16 Mar 2018

Keywords

cell biology
targeted gene modification

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

abstract = "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.",

keywords = "cell biology, targeted gene modification",

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AU - Neguembor, Maria V

AU - Sebastian-Perez, Ruben

AU - Aulicino, Francesco

AU - Gomez-Garcia, Pablo A

AU - Cosma, Maria P

AU - Lakadamyali, Melike

PY - 2018/3/16

Y1 - 2018/3/16

N2 - 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.

AB - 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.

KW - cell biology

KW - targeted gene modification

U2 - 10.1093/nar/gkx1271

DO - 10.1093/nar/gkx1271

M3 - Article (Academic Journal)

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JO - Nucleic Acids Research

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IS - 5

M1 - e30

ER -

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

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