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A Lipid Nanoparticle-Based Method for the Generation of Liver-Specific Knockout Mice
Sumiyo Morita1, Takuro Horii1, Mika Kimura1
1Laboratory of Genome Science, Biosignal Genome Resource Center, Institute for Molecular and Cellular Regulation, Gunma University, 3-39-15 Showa-machi, Maebashi 371-8512, Japan.
Abstract:
Knockout mice are useful tools that can provide information about the normal function of genes, including their biochemical, developmental, and physiological roles. One problem associated with the generation of knockout mice is that the loss of some genes of interest produces a lethal phenotype. Therefore, the use of conditioned knockout mice, in which genes are disrupted in specific organs, is essential for the elucidation of disease pathogenesis and the verification of drug targets. In general, conditional knockout mice are produced using the Cre/loxP system; however, the production of the large numbers of Cre/flox knockout and control mice required for analysis requires substantial time and effort. Here, we describe the generation of liver-specific conditional knockout mice via the introduction of lipid nanoparticles encapsulating Cre mRNA into the liver of floxed mice. This technique does not require the production of offspring by mating floxed mice and is therefore more convenient than the conventional method. The results presented here demonstrate that the LNP-based method enables liver-specific gene knockout in a short period of time.
Insights
Generating conditional knockout mice is crucial for research. A new lipid nanoparticle (LNP) method enables rapid, liver-specific gene knockout in floxed mice, simplifying the process.
Area of Science:
- Genetics and Genomics
- Molecular Biology
- Animal Models
Background:
- Conditional knockout mice are vital for studying gene function and disease pathogenesis.
- Conventional Cre/loxP methods for generating conditional knockout mice are time-consuming and labor-intensive.
- Lethal phenotypes often necessitate conditional gene disruption in specific organs.
Purpose of the Study:
- To develop a more efficient method for generating liver-specific conditional knockout mice.
- To overcome the limitations of traditional breeding strategies for conditional knockout mouse production.
- To facilitate faster research into gene function and drug target validation.
Main Methods:
- Utilized lipid nanoparticles (LNPs) encapsulating Cre mRNA.
- Administered LNPs directly into the liver of floxed mice.
- Bypassed the need for traditional breeding of Cre-driver and floxed mouse lines.
Main Results:
- Achieved efficient and liver-specific gene knockout.
- Demonstrated a significantly reduced timeline for generating knockout mice.
- Confirmed the convenience and efficacy of the LNP-based approach.
Conclusions:
- The LNP-based method offers a rapid and convenient alternative for generating liver-specific conditional knockout mice.
- This technique streamlines the production of essential research models.
- Facilitates accelerated studies in gene function, disease mechanisms, and therapeutic target identification.
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