Establishment of Cre/LoxP-mediated multifunctional reporter knock-in rats with the CRISPR system.

Rats and mice are essential experimental animals in preclinical research, serving as models for various human diseases and contributing significantly to drug development. Rats offer distinct advantages over mice due to their larger size, which allows for more complex surgical procedures, repeated bl...

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Main Authors: Katsuyuki Nakamura, Sara Ito, Yoshihiro Ohguchi, Toshie Jimbo, Yusaku Wada, Ryota Nakajima, Masanobu Kanou, Kei Yamana, Hiroshi Ueda
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2025-01-01
Series:PLoS ONE
Online Access:https://doi.org/10.1371/journal.pone.0325444
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author Katsuyuki Nakamura
Sara Ito
Yoshihiro Ohguchi
Toshie Jimbo
Yusaku Wada
Ryota Nakajima
Masanobu Kanou
Kei Yamana
Hiroshi Ueda
author_facet Katsuyuki Nakamura
Sara Ito
Yoshihiro Ohguchi
Toshie Jimbo
Yusaku Wada
Ryota Nakajima
Masanobu Kanou
Kei Yamana
Hiroshi Ueda
author_sort Katsuyuki Nakamura
collection DOAJ
description Rats and mice are essential experimental animals in preclinical research, serving as models for various human diseases and contributing significantly to drug development. Rats offer distinct advantages over mice due to their larger size, which allows for more complex surgical procedures, repeated blood sampling, or sophisticated behavioral analysis. However, unlike the case with mice, genetically modified rat lines for achieving complex experimental objectives-such as tissue-specific gene knockout or visualization of specific cell populations-are still limited. We here established LoxP-mediated multifunctional reporter KI rats, enabling us to evaluate fluorescence, bioluminescence, and cell-killing assays simultaneously with only one gene-modified rat line. CRISPR/Cas12a, also known as CRISPR/Cpf1, was successfully used to insert the Cre sequence into a target locus to generate Cre driver rats. These results will contribute to the application of gene-modified rats for a more comprehensive understanding of physiology, and for extrapolation of their capabilities in preclinical research.
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institution Kabale University
issn 1932-6203
language English
publishDate 2025-01-01
publisher Public Library of Science (PLoS)
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series PLoS ONE
spelling doaj-art-ecca823e498449f19d54b7eba99322412025-08-20T03:28:10ZengPublic Library of Science (PLoS)PLoS ONE1932-62032025-01-01206e032544410.1371/journal.pone.0325444Establishment of Cre/LoxP-mediated multifunctional reporter knock-in rats with the CRISPR system.Katsuyuki NakamuraSara ItoYoshihiro OhguchiToshie JimboYusaku WadaRyota NakajimaMasanobu KanouKei YamanaHiroshi UedaRats and mice are essential experimental animals in preclinical research, serving as models for various human diseases and contributing significantly to drug development. Rats offer distinct advantages over mice due to their larger size, which allows for more complex surgical procedures, repeated blood sampling, or sophisticated behavioral analysis. However, unlike the case with mice, genetically modified rat lines for achieving complex experimental objectives-such as tissue-specific gene knockout or visualization of specific cell populations-are still limited. We here established LoxP-mediated multifunctional reporter KI rats, enabling us to evaluate fluorescence, bioluminescence, and cell-killing assays simultaneously with only one gene-modified rat line. CRISPR/Cas12a, also known as CRISPR/Cpf1, was successfully used to insert the Cre sequence into a target locus to generate Cre driver rats. These results will contribute to the application of gene-modified rats for a more comprehensive understanding of physiology, and for extrapolation of their capabilities in preclinical research.https://doi.org/10.1371/journal.pone.0325444
spellingShingle Katsuyuki Nakamura
Sara Ito
Yoshihiro Ohguchi
Toshie Jimbo
Yusaku Wada
Ryota Nakajima
Masanobu Kanou
Kei Yamana
Hiroshi Ueda
Establishment of Cre/LoxP-mediated multifunctional reporter knock-in rats with the CRISPR system.
PLoS ONE
title Establishment of Cre/LoxP-mediated multifunctional reporter knock-in rats with the CRISPR system.
title_full Establishment of Cre/LoxP-mediated multifunctional reporter knock-in rats with the CRISPR system.
title_fullStr Establishment of Cre/LoxP-mediated multifunctional reporter knock-in rats with the CRISPR system.
title_full_unstemmed Establishment of Cre/LoxP-mediated multifunctional reporter knock-in rats with the CRISPR system.
title_short Establishment of Cre/LoxP-mediated multifunctional reporter knock-in rats with the CRISPR system.
title_sort establishment of cre loxp mediated multifunctional reporter knock in rats with the crispr system
url https://doi.org/10.1371/journal.pone.0325444
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