Stable Transgenic Mouse Strain with Enhanced Photoactivatable Cre Recombinase for Spatiotemporal Genome Manipulation

Li, Huiying; Ying-yin, WU; Qiu, Yuhao; Li, Xinru; Guan, Yuting; Cao, Xiya; Liu, Meizhen; Zhang, Dan; Huang, Sijie; Lin, Longnian; Hui, Lijian; Ma, Xueyun; Liu, Mingyao; Zhang, Xueli; Wang, Liren; Li, Dali

doi:10.1002/advs.202201352

Cited by 6 publications

(4 citation statements)

References 43 publications

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“…11 ). Indeed, previous research demonstrated that blue light from an LED array can effectively reach mouse liver tissues 31 . Compared to mice injected with the padCas13-2 editor but not exposed to the light stimulation, the RNA editing-enabled mice exhibited a substantially greater luciferase signal (Fig.…”

Section: Resultsmentioning

confidence: 99%

Programmable RNA base editing with photoactivatable CRISPR-Cas13

Yu,

Shin,

et al. 2024

Nat Commun

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CRISPR-Cas13 is widely used for programmable RNA interference, imaging, and editing. In this study, we develop a light-inducible Cas13 system called paCas13 by fusing Magnet with fragment pairs. The most effective split site, N351/C350, was identified and found to exhibit a low background and high inducibility. We observed significant light-induced perturbation of endogenous transcripts by paCas13. We further present a light-inducible base-editing system, herein called the padCas13 editor, by fusing ADAR2 to catalytically inactive paCas13 fragments. The padCas13 editor enabled reversible RNA editing under light and was effective in editing A-to-I and C-to-U RNA bases, targeting disease-relevant transcripts, and fine-tuning endogenous transcripts in mammalian cells in vitro. The padCas13 editor was also used to adjust post-translational modifications and demonstrated the ability to activate target transcripts in a mouse model in vivo. We therefore present a light-inducible RNA-modulating technique based on CRISPR-Cas13 that enables target RNAs to be diversely manipulated in vitro and in vivo, including through RNA degradation and base editing. The approach using the paCas13 system can be broadly applicable to manipulating RNA in various disease states and physiological processes, offering potential additional avenues for research and therapeutic development.

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Section: Resultsmentioning

confidence: 99%

Programmable RNA base editing with photoactivatable CRISPR-Cas13

Yu,

Shin,

et al. 2024

Nat Commun

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“…Over the years, various advancements of the original recombination system have emerged to achieve even finer spatiotemporal comprehension (Shcholok and Eftekharpour, 2023 ). These include the tamoxifen-inducible Cre-ER T2 system (Belteki et al, 2005 ; Heldt and Ressler, 2011 ; Hui et al, 2022 ; Kellogg et al, 2023 ), photoactivatable Cre recombinases (Yoshimi et al, 2021 ; Li et al, 2022 ), Cre/Dre dual recombinase systems (Kouvaros et al, 2023 ) and split Cre systems (Khoo et al, 2020 ; Kim et al, 2021 ), among others. Some of these configurations are of extreme help to easily explore the time and the space, inducing the pathological insult within cell types at various embryonic and postnatal timepoints with high specificity.…”

Section: Exploring Space and Time: Novel Insights From Animal Modelsmentioning

confidence: 99%

Need of orthogonal approaches in neurological disease modeling in mouse

Bossini,

Sessa

2024

Front. Mol. Neurosci.

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Over the years, advancements in modeling neurological diseases have revealed innovative strategies aimed at gaining deeper insights and developing more effective treatments for these complex conditions. However, these progresses have recently been overshadowed by an increasing number of failures in clinical trials, raising doubts about the reliability and translatability of this type of disease modeling. This mini-review does not aim to provide a comprehensive overview of the current state-of-the-art in disease mouse modeling. Instead, it offers a brief excursus over some recent approaches in modeling neurological diseases to pinpoint a few intriguing strategies applied in the field that may serve as sources of inspiration for improving currently available animal models. In particular, we aim to guide the reader toward the potential success of adopting a more orthogonal approach in the study of human diseases.

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“…Optogenetic engineering has been developed to some extent. It has been used for detailed in vivo single-cell analysis because of its efficiency and spatiotemporal specificity ( Yao et al, 2020 ; Li et al, 2022a ; Geiller et al, 2022 ). Here, we describe the advances in cell lineage tracing technology, with a focus on Cre/LoxP recombinase systems ( Figure 1 ).…”

Section: Tracking the Targets All The Way: Lineage Tracingmentioning

confidence: 99%

Adult tissue-specific stem cell interaction: novel technologies and research advances

Luo,

Liu,

2023

Front. Cell Dev. Biol.

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Adult tissue-specific stem cells play a dominant role in tissue homeostasis and regeneration. Various in vivo markers of adult tissue-specific stem cells have been increasingly reported by lineage tracing in genetic mouse models, indicating that marked cells differentiation is crucial during homeostasis and regeneration. How adult tissue-specific stem cells with indicated markers contact the adjacent lineage with indicated markers is of significance to be studied. Novel methods bring future findings. Recent advances in lineage tracing, synthetic receptor systems, proximity labeling, and transcriptomics have enabled easier and more accurate cell behavior visualization and qualitative and quantitative analysis of cell-cell interactions than ever before. These technological innovations have prompted researchers to re-evaluate previous experimental results, providing increasingly compelling experimental results for understanding the mechanisms of cell-cell interactions. This review aimed to describe the recent methodological advances of dual enzyme lineage tracing system, the synthetic receptor system, proximity labeling, single-cell RNA sequencing and spatial transcriptomics in the study of adult tissue-specific stem cells interactions. An enhanced understanding of the mechanisms of adult tissue-specific stem cells interaction is important for tissue regeneration and maintenance of homeostasis in organisms.

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Stable Transgenic Mouse Strain with Enhanced Photoactivatable Cre Recombinase for Spatiotemporal Genome Manipulation

Cited by 6 publications

References 43 publications

Programmable RNA base editing with photoactivatable CRISPR-Cas13

Programmable RNA base editing with photoactivatable CRISPR-Cas13

Need of orthogonal approaches in neurological disease modeling in mouse

Adult tissue-specific stem cell interaction: novel technologies and research advances

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