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Genome editing in the nematode Caenorhabditis briggsae using the CRISPR/Cas9 system

The CRISPR/Cas system has recently emerged as a powerful tool to engineer the genome of an organism. The system is adopted from bacteria where it confers immunity against invading foreign DNA. This work reports the first successful use of the CRISPR/Cas system in Caenorhabditis briggsae (a cousin of...

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Autores principales: Culp, Elizabeth, Richman, Cory, Sharanya, Devika, Jhaveri, Nikita, van den Berg, Wouter, Gupta, Bhagwati P
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7200835/
https://www.ncbi.nlm.nih.gov/pubmed/32395632
http://dx.doi.org/10.1093/biomethods/bpaa003
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author Culp, Elizabeth
Richman, Cory
Sharanya, Devika
Jhaveri, Nikita
van den Berg, Wouter
Gupta, Bhagwati P
author_facet Culp, Elizabeth
Richman, Cory
Sharanya, Devika
Jhaveri, Nikita
van den Berg, Wouter
Gupta, Bhagwati P
author_sort Culp, Elizabeth
collection PubMed
description The CRISPR/Cas system has recently emerged as a powerful tool to engineer the genome of an organism. The system is adopted from bacteria where it confers immunity against invading foreign DNA. This work reports the first successful use of the CRISPR/Cas system in Caenorhabditis briggsae (a cousin of the well-known nematode C. elegans), to generate mutations via non-homologous end joining. We recovered deletion alleles of several conserved genes by microinjecting plasmids that express Cas9 endonuclease and an engineered CRISPR RNA corresponding to the DNA sequence to be cleaved. Evidence for somatic mutations and off-target mutations are also reported. Our approach allows for the generation of loss-of-function mutations in C. briggsae genes thereby facilitating a comparative study of gene function.
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spelling pubmed-72008352020-05-11 Genome editing in the nematode Caenorhabditis briggsae using the CRISPR/Cas9 system Culp, Elizabeth Richman, Cory Sharanya, Devika Jhaveri, Nikita van den Berg, Wouter Gupta, Bhagwati P Biol Methods Protoc Methods Manuscript The CRISPR/Cas system has recently emerged as a powerful tool to engineer the genome of an organism. The system is adopted from bacteria where it confers immunity against invading foreign DNA. This work reports the first successful use of the CRISPR/Cas system in Caenorhabditis briggsae (a cousin of the well-known nematode C. elegans), to generate mutations via non-homologous end joining. We recovered deletion alleles of several conserved genes by microinjecting plasmids that express Cas9 endonuclease and an engineered CRISPR RNA corresponding to the DNA sequence to be cleaved. Evidence for somatic mutations and off-target mutations are also reported. Our approach allows for the generation of loss-of-function mutations in C. briggsae genes thereby facilitating a comparative study of gene function. Oxford University Press 2020-02-10 /pmc/articles/PMC7200835/ /pubmed/32395632 http://dx.doi.org/10.1093/biomethods/bpaa003 Text en © The Author(s) 2020. Published by Oxford University Press. http://creativecommons.org/licenses/by/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Methods Manuscript
Culp, Elizabeth
Richman, Cory
Sharanya, Devika
Jhaveri, Nikita
van den Berg, Wouter
Gupta, Bhagwati P
Genome editing in the nematode Caenorhabditis briggsae using the CRISPR/Cas9 system
title Genome editing in the nematode Caenorhabditis briggsae using the CRISPR/Cas9 system
title_full Genome editing in the nematode Caenorhabditis briggsae using the CRISPR/Cas9 system
title_fullStr Genome editing in the nematode Caenorhabditis briggsae using the CRISPR/Cas9 system
title_full_unstemmed Genome editing in the nematode Caenorhabditis briggsae using the CRISPR/Cas9 system
title_short Genome editing in the nematode Caenorhabditis briggsae using the CRISPR/Cas9 system
title_sort genome editing in the nematode caenorhabditis briggsae using the crispr/cas9 system
topic Methods Manuscript
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7200835/
https://www.ncbi.nlm.nih.gov/pubmed/32395632
http://dx.doi.org/10.1093/biomethods/bpaa003
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