In vivo labeling of endogenous genomic loci in C. elegans using CRISPR/dCas9

Visualization of genomic loci with open chromatin state has been reported in mammalian tissue culture cells using a CRISPR/Cas9-based system that utilizes an EGFP-tagged endonuclease-deficient Cas9 protein (dCas9::EGFP) (Chen et al. 2013). Here, we adapted this approach for use in Caenorhabditis ele...

Descripción completa

Detalles Bibliográficos
Autores principales: Memar, Nadin, Sethi, Aditya, Luehr, Sebastian, Lambie, Eric J, Conradt, Barbara
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Caltech Library 2022
Materias:
New Methods
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9807462/
https://www.ncbi.nlm.nih.gov/pubmed/36606081
http://dx.doi.org/10.17912/micropub.biology.000701
_version_ 1784862724625268736
author Memar, Nadin
Sethi, Aditya
Luehr, Sebastian
Lambie, Eric J
Conradt, Barbara
author_facet Memar, Nadin
Sethi, Aditya
Luehr, Sebastian
Lambie, Eric J
Conradt, Barbara
author_sort Memar, Nadin
collection PubMed
description Visualization of genomic loci with open chromatin state has been reported in mammalian tissue culture cells using a CRISPR/Cas9-based system that utilizes an EGFP-tagged endonuclease-deficient Cas9 protein (dCas9::EGFP) (Chen et al. 2013). Here, we adapted this approach for use in Caenorhabditis elegans . We generated a C. elegans strain that expresses the dCas9 protein fused to two nuclear-localized EGFP molecules (dCas9::NLS::2xEGFP::NLS) in an inducible manner. Using this strain, we report the visualization in live C. elegans embryos of two endogenous repetitive loci, rrn-4 and rrn-1 , from which 5S and 18S ribosomal RNAs are constitutively generated.
format Online
Article
Text
id pubmed-9807462
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher Caltech Library
record_format MEDLINE/PubMed
spelling pubmed-98074622023-01-04 In vivo labeling of endogenous genomic loci in C. elegans using CRISPR/dCas9 Memar, Nadin Sethi, Aditya Luehr, Sebastian Lambie, Eric J Conradt, Barbara MicroPubl Biol New Methods Visualization of genomic loci with open chromatin state has been reported in mammalian tissue culture cells using a CRISPR/Cas9-based system that utilizes an EGFP-tagged endonuclease-deficient Cas9 protein (dCas9::EGFP) (Chen et al. 2013). Here, we adapted this approach for use in Caenorhabditis elegans . We generated a C. elegans strain that expresses the dCas9 protein fused to two nuclear-localized EGFP molecules (dCas9::NLS::2xEGFP::NLS) in an inducible manner. Using this strain, we report the visualization in live C. elegans embryos of two endogenous repetitive loci, rrn-4 and rrn-1 , from which 5S and 18S ribosomal RNAs are constitutively generated. Caltech Library 2022-12-13 /pmc/articles/PMC9807462/ /pubmed/36606081 http://dx.doi.org/10.17912/micropub.biology.000701 Text en Copyright: © 2022 by the authors https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle New Methods
Memar, Nadin
Sethi, Aditya
Luehr, Sebastian
Lambie, Eric J
Conradt, Barbara
In vivo labeling of endogenous genomic loci in C. elegans using CRISPR/dCas9
title In vivo labeling of endogenous genomic loci in C. elegans using CRISPR/dCas9
title_full In vivo labeling of endogenous genomic loci in C. elegans using CRISPR/dCas9
title_fullStr In vivo labeling of endogenous genomic loci in C. elegans using CRISPR/dCas9
title_full_unstemmed In vivo labeling of endogenous genomic loci in C. elegans using CRISPR/dCas9
title_short In vivo labeling of endogenous genomic loci in C. elegans using CRISPR/dCas9
title_sort in vivo labeling of endogenous genomic loci in c. elegans using crispr/dcas9
topic New Methods
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9807462/
https://www.ncbi.nlm.nih.gov/pubmed/36606081
http://dx.doi.org/10.17912/micropub.biology.000701
work_keys_str_mv AT memarnadin invivolabelingofendogenousgenomiclociincelegansusingcrisprdcas9
AT sethiaditya invivolabelingofendogenousgenomiclociincelegansusingcrisprdcas9
AT luehrsebastian invivolabelingofendogenousgenomiclociincelegansusingcrisprdcas9
AT lambieericj invivolabelingofendogenousgenomiclociincelegansusingcrisprdcas9
AT conradtbarbara invivolabelingofendogenousgenomiclociincelegansusingcrisprdcas9

Ejemplares similares