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

Number of Access:778（2025-10-20 13:30 Counts）

Identifier to cite or link to this item: https://hdl.handle.net/11094/100603

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Title
Evaluation of the correlation between nuclear localization levels and genome editing efficiencies of Cas12a fused with nuclear localization signals
Creator
NRID
1000020977342 1000020977342
researchmap permalink
tsukamoto-t tsukamoto-t
CreatorName
Tsukamoto, Tomohito
CreatorName
Mizuta, Haruna
NRID
1000060359859 1000060359859
researchmap permalink
read0068692 read0068692
Scopus Author ID
12647357900 12647357900
CreatorName
Sakai, Eiko
NRID
1000070370939 1000070370939
researchmap permalink
read0165081 read0165081
Scopus Author ID
7003748169 7003748169
CreatorName
Sakurai, Fuminori
NRID
1000050311387 1000050311387
researchmap permalink
read0165079 read0165079
Scopus Author ID
57195381326 57195381326
CreatorName
Mizuguchi, Hiroyuki
Subject
Adenovirus vector
Cas12a
Gene therapy
Genome editing
Nuclear localization signal
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.
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.
Publisher
Elsevier B.V.
Source Title
Journal of Pharmaceutical Sciences
Volume (Issue)
114(2)
Page
841-848
Date of Issued
2025-02-01
Language
English
Handle URL
https://hdl.handle.net/11094/100603
PISSN
00223549
eISSN
15206017
Relation.isIdenticalTo
DOI (Publisher Version)
https://doi.org/10.1016/j.xphs.2024.10.029
Access Rights
open access
CopyRight
This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
oaire:version
Version of Record
Category
学術雑誌論文 Journal Article