Evaluation of the correlation between nuclear localization levels and genome editing efficiencies of Cas12a fused with nuclear localization signals

Tsukamoto, Tomohito; Mizuta, Haruna; Sakai, Eiko; Sakurai, Fuminori; Mizuguchi, Hiroyuki

doi:https://doi.org/10.1016/j.xphs.2024.10.029

Evaluation of the correlation between nuclear localization levels and genome editing efficiencies of Cas12a fused with nuclear localization signals

Tsukamoto, Tomohito; Mizuta, Haruna; Sakai, Eiko et al.

Journal of Pharmaceutical Sciences, 2025, 114(2), 841-848

アクセス数:748件（2025-09-25 14:39 集計）

固定URL: https://hdl.handle.net/11094/100603

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タイトル	Evaluation of the correlation between nuclear localization levels and genome editing efficiencies of Cas12a fused with nuclear localization signals
著者	NRID 1000020977342 1000020977342 researchmap permalink tsukamoto-t tsukamoto-t 著者名 Tsukamoto, Tomohito
	著者名 Mizuta, Haruna
	NRID 1000060359859 1000060359859 researchmap permalink read0068692 read0068692 Scopus Author ID 12647357900 12647357900 著者名 Sakai, Eiko
	NRID 1000070370939 1000070370939 researchmap permalink read0165081 read0165081 Scopus Author ID 7003748169 7003748169 著者名 Sakurai, Fuminori
	NRID 1000050311387 1000050311387 researchmap permalink read0165079 read0165079 Scopus Author ID 57195381326 57195381326 著者名 Mizuguchi, Hiroyuki
キーワード等	Adenovirus vector
	Cas12a
	Gene therapy
	Genome editing
	Nuclear localization signal
内容	Tsukamoto T., Mizuta H., Sakai E., et al. Evaluation of the correlation between nuclear localization levels and genome editing efficiencies of Cas12a fused with nuclear localization signals. Journal of Pharmaceutical Sciences 114, 841 (2025); https://doi.org/10.1016/j.xphs.2024.10.029.
抄録	Genome editing technology using the CRISPR-Cas system is attracting much attention not only as a promising experimental tool for analysis of genome functions, but also as a novel therapeutic approach for genetic disorders. Among the various types of Cas proteins, Cas12a is expected to be a promising gene editing tool due to its unique properties, including low off-target effects. As Cas proteins are of prokaryotic origin, they need to be fused with appropriate localization signals to perform their function in eukaryotic cells. Cas12a proteins fused with a nuclear localization signal (NLS) have been developed so far, but the relation between the nuclear localization activity and the genome editing efficiency has not been fully elucidated. Here, utilizing two Cas12a orthologs, AsCas12a and LbCas12a, with various number of NLSs derived from various origins, we revealed that the improved nuclear localization resulted in increased genome editing efficiencies when expressed using adenovirus (Ad) vector in cultured cells. However, when they were expressed in mouse liver, the improvement of the nuclear localization activity was not necessarily required to achieve the maximum genome editing efficiency four weeks after Ad vector administration. These data indicated that the optimized NLS modification of Cas12a proteins in vitro situations differed from that in vivo.
出版者	Elsevier B.V.
収録物名	Journal of Pharmaceutical Sciences
巻(号)	114(2)
ページ	841-848
刊行年月	2025-02-01
言語	英語
ハンドルURL	https://hdl.handle.net/11094/100603
PISSN	00223549
eISSN	15206017
関連情報 (isIdenticalTo)	DOI (出版社版) https://doi.org/10.1016/j.xphs.2024.10.029
アクセス権	open access
権利情報	This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 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
DCTERMS.bibliographicCitation	Journal of Pharmaceutical Sciences. 2025, 114(2), p. 841-848
DC.title	Evaluation of the correlation between nuclear localization levels and genome editing efficiencies of Cas12a fused with nuclear localization signals
DC.creator	Tsukamoto, Tomohito
	Mizuta, Haruna
	Sakai, Eiko
	Sakurai, Fuminori
	Mizuguchi, Hiroyuki
DC.publisher	Elsevier B.V.
DC.language' scheme='DCTERMS.RFC1766	英語
DCTERMS.issued' scheme='DCTERMS.W3CDTF	2025-02-01
DC.identifier	https://doi.org/10.1016/j.xphs.2024.10.029
DC.subject	Adenovirus vector
	Cas12a
	Gene therapy
	Genome editing
	Nuclear localization signal
DC.description	Tsukamoto T., Mizuta H., Sakai E., et al. Evaluation of the correlation between nuclear localization levels and genome editing efficiencies of Cas12a fused with nuclear localization signals. Journal of Pharmaceutical Sciences 114, 841 (2025); https://doi.org/10.1016/j.xphs.2024.10.029.
DCTERMS.abstract	Genome editing technology using the CRISPR-Cas system is attracting much attention not only as a promising experimental tool for analysis of genome functions, but also as a novel therapeutic approach for genetic disorders. Among the various types of Cas proteins, Cas12a is expected to be a promising gene editing tool due to its unique properties, including low off-target effects. As Cas proteins are of prokaryotic origin, they need to be fused with appropriate localization signals to perform their function in eukaryotic cells. Cas12a proteins fused with a nuclear localization signal (NLS) have been developed so far, but the relation between the nuclear localization activity and the genome editing efficiency has not been fully elucidated. Here, utilizing two Cas12a orthologs, AsCas12a and LbCas12a, with various number of NLSs derived from various origins, we revealed that the improved nuclear localization resulted in increased genome editing efficiencies when expressed using adenovirus (Ad) vector in cultured cells. However, when they were expressed in mouse liver, the improvement of the nuclear localization activity was not necessarily required to achieve the maximum genome editing efficiency four weeks after Ad vector administration. These data indicated that the optimized NLS modification of Cas12a proteins in vitro situations differed from that in vivo.
DC.rights	This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
DC.format	100603
citation_title	Evaluation of the correlation between nuclear localization levels and genome editing efficiencies of Cas12a fused with nuclear localization signals
citation_author	Tsukamoto, Tomohito
	Mizuta, Haruna
	Sakai, Eiko
	Sakurai, Fuminori
	Mizuguchi, Hiroyuki
citation_publisher	Elsevier B.V.
citation_language	英語
citation_date	2025-02-01
citation_journal_title	Journal of Pharmaceutical Sciences
citation_volume	114
citation_issue	2
citation_firstpage	841
citation_lastpage	848
citation_doi	https://doi.org/10.1016/j.xphs.2024.10.029
citation_public_url	https://hdl.handle.net/11094/100603

Evaluation of the correlation between nuclear localization levels and genome editing efficiencies of Cas12a fused with nuclear localization signals

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