A blueprint for gene function analysis through Base Editing in the model plant Physcomitrium (Physcomitrella) patens

CRISPR-Cas9 has proven to be highly valuable for genome editing in plants, including the model plant Physcomitrium patens. However, the fact that most of the editing events produced using the native Cas9 nuclease correspond to small insertions and deletions is a limitation. CRISPR-Cas9 base editors...

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Autores: Guyon-Debast, Anouchka|||0000-0003-3489-1035, Alboresi, Alessandro|||0000-0003-4818-7778, Terret, Zoé|||0000-0002-8057-755X, Charlot, Florence|||0000-0003-1218-951X, Berthier, Floriane|||0000-0003-3368-8078, Vendrell Mir, Pol|||0000-0002-5695-5880, Casacuberta, Josep M.|||0000-0002-5609-4152, Veillet, Florian|||0000-0002-6892-6825, Morosinotto, Tomas|||0000-0002-0803-7591, Gallois, Jean-Luc|||0000-0003-0451-1740, Nogué, Fabien|||0000-0003-0619-4638
Tipo de recurso: artículo
Fecha de publicación:2021
País:España
Institución:Universitat Autònoma de Barcelona
Repositorio:Dipòsit Digital de Documents de la UAB
Idioma:inglés
OAI Identifier:oai:ddd.uab.cat:252290
Acceso en línea:https://ddd.uab.cat/record/252290
https://dx.doi.org/urn:doi:10.1111/nph.17171
Access Level:acceso abierto
Palabra clave:Adenine deaminase
APRT
Base editing
Cas9
CRISPR
Cytosine deaminase
Physcomitrella patens
Physcomitrium patens
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repository_id_str
spelling A blueprint for gene function analysis through Base Editing in the model plant Physcomitrium (Physcomitrella) patensGuyon-Debast, Anouchka|||0000-0003-3489-1035Alboresi, Alessandro|||0000-0003-4818-7778Terret, Zoé|||0000-0002-8057-755XCharlot, Florence|||0000-0003-1218-951XBerthier, Floriane|||0000-0003-3368-8078Vendrell Mir, Pol|||0000-0002-5695-5880Casacuberta, Josep M.|||0000-0002-5609-4152Veillet, Florian|||0000-0002-6892-6825Morosinotto, Tomas|||0000-0002-0803-7591Gallois, Jean-Luc|||0000-0003-0451-1740Nogué, Fabien|||0000-0003-0619-4638Adenine deaminaseAPRTBase editingCas9CRISPRCytosine deaminasePhyscomitrella patensPhyscomitrium patensCRISPR-Cas9 has proven to be highly valuable for genome editing in plants, including the model plant Physcomitrium patens. However, the fact that most of the editing events produced using the native Cas9 nuclease correspond to small insertions and deletions is a limitation. CRISPR-Cas9 base editors enable targeted mutation of single nucleotides in eukaryotic genomes and therefore overcome this limitation. Here, we report two programmable base-editing systems to induce precise cytosine or adenine conversions in P. patens. Using cytosine or adenine base editors, site-specific single-base mutations can be achieved with an efficiency up to 55%, without off-target mutations. Using the APT gene as a reporter of editing, we could show that both base editors can be used in simplex or multiplex, allowing for the production of protein variants with multiple amino-acid changes. Finally, we set up a co-editing selection system, named selecting modification of APRT to report gene targeting (SMART), allowing up to 90% efficiency site-specific base editing in P. patens. These two base editors will facilitate gene functional analysis in P. patens, allowing for site-specific editing of a given base through single sgRNA base editing or for in planta evolution of a given gene through the production of randomly mutagenised variants using multiple sgRNA base editing. 22021-01-0120212021-01-01Articlehttp://purl.org/coar/resource_type/c_6501VoRhttp://purl.org/coar/version/c_970fb48d4fbd8a85info:eu-repo/semantics/articleapplication/pdfhttps://ddd.uab.cat/record/252290https://dx.doi.org/urn:doi:10.1111/nph.17171reponame:Dipòsit Digital de Documents de la UABinstname:Universitat Autònoma de BarcelonaInglésengopen accesshttp://purl.org/coar/access_right/c_abf2Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, i la comunicació pública de l'obra, sempre que no sigui amb finalitats comercials, i sempre que es reconegui l'autoria de l'obra original. No es permet la creació d'obres derivades.https://creativecommons.org/licenses/by-nc-nd/4.0/info:eu-repo/semantics/openAccessoai:ddd.uab.cat:2522902026-06-06T12:50:31Z
dc.title.none.fl_str_mv A blueprint for gene function analysis through Base Editing in the model plant Physcomitrium (Physcomitrella) patens
title A blueprint for gene function analysis through Base Editing in the model plant Physcomitrium (Physcomitrella) patens
spellingShingle A blueprint for gene function analysis through Base Editing in the model plant Physcomitrium (Physcomitrella) patens
Guyon-Debast, Anouchka|||0000-0003-3489-1035
Adenine deaminase
APRT
Base editing
Cas9
CRISPR
Cytosine deaminase
Physcomitrella patens
Physcomitrium patens
title_short A blueprint for gene function analysis through Base Editing in the model plant Physcomitrium (Physcomitrella) patens
title_full A blueprint for gene function analysis through Base Editing in the model plant Physcomitrium (Physcomitrella) patens
title_fullStr A blueprint for gene function analysis through Base Editing in the model plant Physcomitrium (Physcomitrella) patens
title_full_unstemmed A blueprint for gene function analysis through Base Editing in the model plant Physcomitrium (Physcomitrella) patens
title_sort A blueprint for gene function analysis through Base Editing in the model plant Physcomitrium (Physcomitrella) patens
dc.creator.none.fl_str_mv Guyon-Debast, Anouchka|||0000-0003-3489-1035
Alboresi, Alessandro|||0000-0003-4818-7778
Terret, Zoé|||0000-0002-8057-755X
Charlot, Florence|||0000-0003-1218-951X
Berthier, Floriane|||0000-0003-3368-8078
Vendrell Mir, Pol|||0000-0002-5695-5880
Casacuberta, Josep M.|||0000-0002-5609-4152
Veillet, Florian|||0000-0002-6892-6825
Morosinotto, Tomas|||0000-0002-0803-7591
Gallois, Jean-Luc|||0000-0003-0451-1740
Nogué, Fabien|||0000-0003-0619-4638
author Guyon-Debast, Anouchka|||0000-0003-3489-1035
author_facet Guyon-Debast, Anouchka|||0000-0003-3489-1035
Alboresi, Alessandro|||0000-0003-4818-7778
Terret, Zoé|||0000-0002-8057-755X
Charlot, Florence|||0000-0003-1218-951X
Berthier, Floriane|||0000-0003-3368-8078
Vendrell Mir, Pol|||0000-0002-5695-5880
Casacuberta, Josep M.|||0000-0002-5609-4152
Veillet, Florian|||0000-0002-6892-6825
Morosinotto, Tomas|||0000-0002-0803-7591
Gallois, Jean-Luc|||0000-0003-0451-1740
Nogué, Fabien|||0000-0003-0619-4638
author_role author
author2 Alboresi, Alessandro|||0000-0003-4818-7778
Terret, Zoé|||0000-0002-8057-755X
Charlot, Florence|||0000-0003-1218-951X
Berthier, Floriane|||0000-0003-3368-8078
Vendrell Mir, Pol|||0000-0002-5695-5880
Casacuberta, Josep M.|||0000-0002-5609-4152
Veillet, Florian|||0000-0002-6892-6825
Morosinotto, Tomas|||0000-0002-0803-7591
Gallois, Jean-Luc|||0000-0003-0451-1740
Nogué, Fabien|||0000-0003-0619-4638
author2_role author
author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Adenine deaminase
APRT
Base editing
Cas9
CRISPR
Cytosine deaminase
Physcomitrella patens
Physcomitrium patens
topic Adenine deaminase
APRT
Base editing
Cas9
CRISPR
Cytosine deaminase
Physcomitrella patens
Physcomitrium patens
description CRISPR-Cas9 has proven to be highly valuable for genome editing in plants, including the model plant Physcomitrium patens. However, the fact that most of the editing events produced using the native Cas9 nuclease correspond to small insertions and deletions is a limitation. CRISPR-Cas9 base editors enable targeted mutation of single nucleotides in eukaryotic genomes and therefore overcome this limitation. Here, we report two programmable base-editing systems to induce precise cytosine or adenine conversions in P. patens. Using cytosine or adenine base editors, site-specific single-base mutations can be achieved with an efficiency up to 55%, without off-target mutations. Using the APT gene as a reporter of editing, we could show that both base editors can be used in simplex or multiplex, allowing for the production of protein variants with multiple amino-acid changes. Finally, we set up a co-editing selection system, named selecting modification of APRT to report gene targeting (SMART), allowing up to 90% efficiency site-specific base editing in P. patens. These two base editors will facilitate gene functional analysis in P. patens, allowing for site-specific editing of a given base through single sgRNA base editing or for in planta evolution of a given gene through the production of randomly mutagenised variants using multiple sgRNA base editing.
publishDate 2021
dc.date.none.fl_str_mv 2
2021-01-01
2021
2021-01-01
dc.type.none.fl_str_mv Article
http://purl.org/coar/resource_type/c_6501
VoR
http://purl.org/coar/version/c_970fb48d4fbd8a85
dc.type.openaire.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.none.fl_str_mv https://ddd.uab.cat/record/252290
https://dx.doi.org/urn:doi:10.1111/nph.17171
url https://ddd.uab.cat/record/252290
https://dx.doi.org/urn:doi:10.1111/nph.17171
dc.language.none.fl_str_mv Inglés
eng
language_invalid_str_mv Inglés
language eng
dc.rights.none.fl_str_mv open access
http://purl.org/coar/access_right/c_abf2
https://creativecommons.org/licenses/by-nc-nd/4.0/
dc.rights.openaire.fl_str_mv info:eu-repo/semantics/openAccess
rights_invalid_str_mv open access
http://purl.org/coar/access_right/c_abf2
https://creativecommons.org/licenses/by-nc-nd/4.0/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.source.none.fl_str_mv reponame:Dipòsit Digital de Documents de la UAB
instname:Universitat Autònoma de Barcelona
instname_str Universitat Autònoma de Barcelona
reponame_str Dipòsit Digital de Documents de la UAB
collection Dipòsit Digital de Documents de la UAB
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15,300719

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