A Ratiometric Gene-Switch System for miRNA Sensing and Gene Regulation.
cancer therapy
gene switch
miRNA imaging
transcriptional repressor
Journal
Small methods
ISSN: 2366-9608
Titre abrégé: Small Methods
Pays: Germany
ID NLM: 101724536
Informations de publication
Date de publication:
Mar 2024
Mar 2024
Historique:
revised:
28
10
2023
received:
19
09
2023
pubmed:
27
11
2023
medline:
27
11
2023
entrez:
27
11
2023
Statut:
ppublish
Résumé
microRNAs (miRNAs) are a class of non-coding, small RNAs that play an important role in diverse biological processes and diseases. By regulating the expression of eukaryotic genes post-transcriptionally in a sequence-specific manner, miRNAs are widely used to design synthetic RNA switches. However, most of the RNA switches are often dependent on the corresponding ligand molecules, whose specificity and concentration would affect the efficiency of synthetic RNA circuits. Here, a fused transcriptional repressor Gal4BD-Rluc based gene-switch system Gal-miR for miRNA visualization and gene regulation is described. By placing a luciferase downstream gene under the control of endogenous miRNA machinery, the Gal-miR system makes the conversion of miRNA-mediated gene silencing into a ratiometric bioluminescent signal, which quantitatively reflected miRNA-206 activity during myogenic differentiation. Moreover, it demonstrates that this gene-switch system can effectively inhibit breast cancer cell viability, migration and invasion under the control of specific miRNAs by replacing the downstream gene with melittin functional gene. The study proposes a powerful modular genetic design for achieving precise control of transgene expression in a miRNA responsive way, as well as visualizing the dynamics of miRNA activity.
Identifiants
pubmed: 38009771
doi: 10.1002/smtd.202301266
doi:
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
e2301266Subventions
Organisme : National Natural Science Foundation of China
ID : 32271512
Organisme : Natural Science Basic Research Program of Shaanxi
ID : 2022JC-56
Organisme : Natural Science Basic Research Program of Shaanxi
ID : 2023-JC-ZD-43
Organisme : Natural Science Basic Research Program of Shaanxi
ID : 2023-JC-YB-660
Organisme : Natural Science Basic Research Program of Shaanxi
ID : 2022JM-468
Organisme : Foreign Expert Project of Ministry of Science and Technology of China
ID : G2023174001L
Organisme : Shaanxi Fundamental Science Research Project for Chemistry & Biology
ID : 22JHQ088
Organisme : Social Talent Grant of Tangdu Hospital of Air Force Medical University
ID : 2021SHRC026
Informations de copyright
© 2023 Wiley-VCH GmbH.
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