Development of programmable RNA imaging with RNA-guided GFP via click chemistry

Nakamura, Jun; Shiraishi, Miyako; Yamamoto, Junpei; Suzuki, Keiichiro

doi:https://doi.org/10.1093/nar/gkaf1147

Development of programmable RNA imaging with RNA-guided GFP via click chemistry

Nakamura, Jun; Shiraishi, Miyako; Yamamoto, Junpei et al.

Nucleic Acids Research, 2025, 53(20), gkaf1147

アクセス数:40件（2026-02-16 01:33 集計）

固定URL: https://hdl.handle.net/11094/103679

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タイトル	Development of programmable RNA imaging with RNA-guided GFP via click chemistry
著者	著者名 Nakamura, Jun
	NRID 1000000813940 1000000813940 著者名 Shiraishi, Miyako
	NRID 1000090571084 1000090571084 ORCID 0000-0002-5036-991X 0000-0002-5036-991X researchmap permalink yamamotojunpei yamamotojunpei Scopus Author ID 15019956000 15019956000 著者名 Yamamoto, Junpei
	NRID 1000070433654 1000070433654 ORCID 0000-0001-5603-9105 0000-0001-5603-9105 researchmap permalink ksuzuki ksuzuki Scopus Author ID 55389105500 55389105500 著者名 Suzuki, Keiichiro
内容	Nakamura J., Shiraishi M., Yamamoto J., et al. Development of programmable RNA imaging with RNA-guided GFP via click chemistry. Nucleic Acids Research 53, gkaf1147 (2025); https://doi.org/10.1093/nar/gkaf1147.
抄録	The CRISPR-Cas system revolutionized molecular biology by guiding Cas proteins to target nucleic acid sequences using customizable guide RNAs, offering unparalleled precision and versatility. Inspired by this innovation, we developed RNA-guided green fluorescent protein (RGG), a simple and programmable platform for targeting nucleic acid. Using a streamlined click chemistry approach, known for its high efficiency and specificity, we conjugated dibenzocyclooctyne (DBCO)-modified guide nucleic acids, designed to complement target sequences, with azide-exposed proteins to construct RGG. Systematic optimization identified 30-nt RNA with 3′-DBCO modifications as the most effective configuration for RGG, enabling precise visualization of nuclear-localized RNAs, including NEAT1 and Satellite III RNA, in living cells. This establishes RGG as a customizable and efficient system for RNA imaging and molecular analysis, underscoring the potential of direct conjugation between guide nucleic acids and proteins to enable precise nucleic acid recognition and dynamic molecular modification in living cells.
出版者	Oxford University Press
収録物名	Nucleic Acids Research
巻(号)	53(20)
ページ	gkaf1147
刊行年月	2025-11-08
言語	英語
ハンドルURL	https://hdl.handle.net/11094/103679
PISSN	03051048
eISSN	13624962
関連情報 (isIdenticalTo)	DOI (出版社版) https://doi.org/10.1093/nar/gkaf1147
アクセス権	open access
権利情報	This article is licensed under a Creative Commons Attribution 4.0 International License.
出版タイプ	Version of Record
カテゴリ	学術雑誌論文 Journal Article

資源タイプ	学術雑誌論文
ローカル資源タイプ	学術雑誌論文
DCMI資源タイプ	text
DCTERMS.bibliographicCitation	Nucleic Acids Research. 2025, 53(20), p. gkaf1147
DC.title	Development of programmable RNA imaging with RNA-guided GFP via click chemistry
DC.creator	Nakamura, Jun
	Shiraishi, Miyako
	Yamamoto, Junpei
	Suzuki, Keiichiro
DC.publisher	Oxford University Press
DC.language' scheme='DCTERMS.RFC1766	英語
DC.type' scheme='DCTERMS.DCMIType	text
DCTERMS.issued' scheme='DCTERMS.W3CDTF	2025-11-08
DC.identifier	https://doi.org/10.1093/nar/gkaf1147
DC.description	Nakamura J., Shiraishi M., Yamamoto J., et al. Development of programmable RNA imaging with RNA-guided GFP via click chemistry. Nucleic Acids Research 53, gkaf1147 (2025); https://doi.org/10.1093/nar/gkaf1147.
DCTERMS.abstract	The CRISPR-Cas system revolutionized molecular biology by guiding Cas proteins to target nucleic acid sequences using customizable guide RNAs, offering unparalleled precision and versatility. Inspired by this innovation, we developed RNA-guided green fluorescent protein (RGG), a simple and programmable platform for targeting nucleic acid. Using a streamlined click chemistry approach, known for its high efficiency and specificity, we conjugated dibenzocyclooctyne (DBCO)-modified guide nucleic acids, designed to complement target sequences, with azide-exposed proteins to construct RGG. Systematic optimization identified 30-nt RNA with 3′-DBCO modifications as the most effective configuration for RGG, enabling precise visualization of nuclear-localized RNAs, including NEAT1 and Satellite III RNA, in living cells. This establishes RGG as a customizable and efficient system for RNA imaging and molecular analysis, underscoring the potential of direct conjugation between guide nucleic acids and proteins to enable precise nucleic acid recognition and dynamic molecular modification in living cells.
DC.rights	This article is licensed under a Creative Commons Attribution 4.0 International License.
DC.format	103679
citation_title	Development of programmable RNA imaging with RNA-guided GFP via click chemistry
citation_author	Nakamura, Jun
	Shiraishi, Miyako
	Yamamoto, Junpei
	Suzuki, Keiichiro
citation_publisher	Oxford University Press
citation_language	英語
citation_date	2025-11-08
citation_journal_title	Nucleic Acids Research
citation_volume	53
citation_issue	20
citation_firstpage	gkaf1147
citation_doi	https://doi.org/10.1093/nar/gkaf1147
citation_public_url	https://hdl.handle.net/11094/103679

Development of programmable RNA imaging with RNA-guided GFP via click chemistry

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