PUBLICATION

Generation of ints14 Knockout Zebrafish using CRISPR/Cas9 for the Study of Development and Disease Mechanisms

Authors
Jung, J.H., Jeon, S., Kim, H., Jung, S.H.
ID
ZDB-PUB-240131-12
Date
2023
Source
Development & reproduction   27: 205211205-211 (Journal)
Registered Authors
Jung, Seung-Hyun
Keywords
CRISPR/Cas9, INTS14, Mutation, RNA polymerase II, Zebrafish
MeSH Terms
none
PubMed
38292235 Full text @ Dev Reprod
Abstract
INTS14/VWA9, a component of the integrator complex subunits, plays a pivotal role in regulating the fate of numerous nascent RNAs transcribed by RNA polymerase II, particularly in the biogenesis of small nuclear RNAs and enhancer RNAs. Despite its significance, a comprehensive mutation model for developmental research has been lacking. To address this gap, we aimed to investigate the expression patterns of INTS14 during zebrafish embryonic development. We generated ints14 mutant strains using the CRISPR/Cas9 system. We validated the gRNA activity by co-injecting Cas9 protein and a single guide RNA into fertilized zebrafish eggs, subsequently confirming the presence of a 6- or 9-bp deletion in the ints14 gene. In addition, we examined the two mutant alleles through PCR analysis, T7E1 assay, TA-cloning, and sequencing. For the first time, we used the CRISPR/Cas9 system to create a model in which some sequences of the ints14 gene were removed. This breakthrough opens new avenues for in-depth exploration of the role of ints14 in animal diseases. The mutant strains generated in this study can provide a valuable resource for further investigations into the specific consequences of ints14 gene deletion during zebrafish development. This research establishes a foundation for future studies exploring the molecular mechanisms underlying the functions of ints14, its interactions with other genes or proteins, and its broader implications for biological processes.
Genes / Markers
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Expression
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Phenotype
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Mutations / Transgenics
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Human Disease / Model
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Sequence Targeting Reagents
Target Reagent Reagent Type
ints14CRISPR1-ints14CRISPR
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Fish
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Antibodies
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Orthology
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Engineered Foreign Genes
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