Establishment of CRISPR/Cas9-based knock-in in a hemimetabolous insect : targeted gene tagging in the cricket Gryllus bimaculatus

2025
journal article
article
dc.abstract.enStudies of traditional model organisms like the fruit fly Drosophila melanogaster have contributed immensely to our understanding of the genetic basis of developmental processes. However, the generalizability of these findings cannot be confirmed without functional genetic analyses in additional organisms. Direct genome editing using targeted nucleases has the potential to transform hitherto poorly-understood organisms into viable laboratory organisms for functional genetic study. To this end, here we present a method to induce targeted genome knock-out and knock-in of desired sequences in an insect that serves as an informative contrast to Drosophila, the cricket Gryllus bimaculatus. The efficiency of germ line transmission of induced mutations is comparable to that reported for other well-studied laboratory organisms, and knock-ins targeting introns yield viable, fertile animals in which knock-in events are directly detectable by visualization of a fluorescent marker in the expression pattern of the targeted gene. Combined with the recently assembled and annotated genome of this cricket, this knock-in/knock-out method increases the viability of G. bimaculatus as a tractable system for functional genetics in a basally branching insect.
dc.affiliationWydział Biochemii, Biofizyki i Biotechnologii : Pracownia Bioinformatyki i Biologii Genomu
dc.contributor.authorMatsuoka, Yuji
dc.contributor.authorNakamura, Taro
dc.contributor.authorWatanabe, Takahito
dc.contributor.authorBarnett, Austen A.
dc.contributor.authorTomonari, Sayuri
dc.contributor.authorYlla Bou, Guillem - 460894
dc.contributor.authorWhittle, Carrie A.
dc.contributor.authorNoji, Sumihare
dc.contributor.authorMito, Taro
dc.contributor.authorExtavour, Cassandra G.
dc.date.accessioned2025-01-13T09:16:10Z
dc.date.available2025-01-13T09:16:10Z
dc.date.createdat2025-01-13T09:16:10Zen
dc.date.issued2025
dc.date.openaccess0
dc.description.accesstimew momencie opublikowania
dc.description.additionalGuillem Ylla Bou podpisany: Guillem Ylla. Bibliogr.
dc.description.number1
dc.description.versionostateczna wersja wydawcy
dc.description.volume152
dc.identifier.articleiddev199746
dc.identifier.doi10.1242/dev.199746
dc.identifier.eissn1477-9129
dc.identifier.issn0950-1991
dc.identifier.urihttps://ruj.uj.edu.pl/handle/item/539747
dc.languageeng
dc.language.containereng
dc.rightsUdzielam licencji. Uznanie autorstwa 4.0 Międzynarodowa
dc.rights.licenceCC-BY
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/legalcode.pl
dc.share.typeinne
dc.subject.enCRISPR/Cas9
dc.subject.enOrthoptera
dc.subject.engenome editing
dc.subject.enHox genes
dc.subject.enUltrabithorax
dc.subject.enabdominal-A
dc.subtypeArticle
dc.titleEstablishment of CRISPR/Cas9-based knock-in in a hemimetabolous insect : targeted gene tagging in the cricket Gryllus bimaculatus
dc.title.journalDevelopment (Cambridge)
dc.typeJournalArticle
dspace.entity.typePublicationen
dc.abstract.en
Studies of traditional model organisms like the fruit fly Drosophila melanogaster have contributed immensely to our understanding of the genetic basis of developmental processes. However, the generalizability of these findings cannot be confirmed without functional genetic analyses in additional organisms. Direct genome editing using targeted nucleases has the potential to transform hitherto poorly-understood organisms into viable laboratory organisms for functional genetic study. To this end, here we present a method to induce targeted genome knock-out and knock-in of desired sequences in an insect that serves as an informative contrast to Drosophila, the cricket Gryllus bimaculatus. The efficiency of germ line transmission of induced mutations is comparable to that reported for other well-studied laboratory organisms, and knock-ins targeting introns yield viable, fertile animals in which knock-in events are directly detectable by visualization of a fluorescent marker in the expression pattern of the targeted gene. Combined with the recently assembled and annotated genome of this cricket, this knock-in/knock-out method increases the viability of G. bimaculatus as a tractable system for functional genetics in a basally branching insect.
dc.affiliation
Wydział Biochemii, Biofizyki i Biotechnologii : Pracownia Bioinformatyki i Biologii Genomu
dc.contributor.author
Matsuoka, Yuji
dc.contributor.author
Nakamura, Taro
dc.contributor.author
Watanabe, Takahito
dc.contributor.author
Barnett, Austen A.
dc.contributor.author
Tomonari, Sayuri
dc.contributor.author
Ylla Bou, Guillem - 460894
dc.contributor.author
Whittle, Carrie A.
dc.contributor.author
Noji, Sumihare
dc.contributor.author
Mito, Taro
dc.contributor.author
Extavour, Cassandra G.
dc.date.accessioned
2025-01-13T09:16:10Z
dc.date.available
2025-01-13T09:16:10Z
dc.date.createdaten
2025-01-13T09:16:10Z
dc.date.issued
2025
dc.date.openaccess
0
dc.description.accesstime
w momencie opublikowania
dc.description.additional
Guillem Ylla Bou podpisany: Guillem Ylla. Bibliogr.
dc.description.number
1
dc.description.version
ostateczna wersja wydawcy
dc.description.volume
152
dc.identifier.articleid
dev199746
dc.identifier.doi
10.1242/dev.199746
dc.identifier.eissn
1477-9129
dc.identifier.issn
0950-1991
dc.identifier.uri
https://ruj.uj.edu.pl/handle/item/539747
dc.language
eng
dc.language.container
eng
dc.rights
Udzielam licencji. Uznanie autorstwa 4.0 Międzynarodowa
dc.rights.licence
CC-BY
dc.rights.uri
http://creativecommons.org/licenses/by/4.0/legalcode.pl
dc.share.type
inne
dc.subject.en
CRISPR/Cas9
dc.subject.en
Orthoptera
dc.subject.en
genome editing
dc.subject.en
Hox genes
dc.subject.en
Ultrabithorax
dc.subject.en
abdominal-A
dc.subtype
Article
dc.title
Establishment of CRISPR/Cas9-based knock-in in a hemimetabolous insect : targeted gene tagging in the cricket Gryllus bimaculatus
dc.title.journal
Development (Cambridge)
dc.type
JournalArticle
dspace.entity.typeen
Publication
Affiliations

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