Myo1h基因敲除小鼠模型的建立与初步观察

doi:10.13591/j.cnki.kqyx.2022.08.001

Abstract

Abstract: Objective To construct Myo1h gene homologous to human exon 30 knockout model mice (Myo1h mice) and to verify its knockout efficiency. Methods The F0 generation of Myo1h knockout mice was established with Crispr/Cas9 technology and homozygous F2 generation mice were obtained after breeding and genotype identification. Knockout efficiency at mRNA level and protein level was validated by real-time fluorescence quantitative PCR and Western Blot. Immunofluorescence assay was used to observe the expression of Myo1h in the condyle of mice to verify the knockout efficiency in vivo. Phenotypic assessment of F2 generation mice was carried out by measuring effective mandibular length through Micro-CT 3D reconstruction. Prism 8.0 software package was used for statistical analysis. Results Real-time quantitative PCR and western blot results showed that the mRNA level of Myo1h gene was significantly decreased (P<0.05), and protein expression was reduced. Immunofluorescence assay results showed that the expression of Myo1h in brain, lung tissues and condyle of Myo1h mice was significantly decreased (P<0.05). Phenotypic assessment by Micro-CT and length measurement analysis revealed that the effective body length of Myo1h knockout mice was shorter than that of control group (P<0.05). Conclusion In this study, model of Myo1h gene knockout mice is successfully constructed and the body length development of knockout mice is inhibited compared to control group. With further study on developmental effects of the Myo1h gene, it may provide new ideas for study on cranio-maxillofacial bone growth and development.

Key words: Myo1h gene, mouse, knock-out, genotyping, growth and development

CLC Number:

R349.64

WEN Shiqiang, SUN Rongrong, SHI Han, LI Yongming. Construction and observation of Myo1h gene knockout mouse model[J]. Stomatology, 2022, 42(8): 673-680.

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Construction and observation of Myo1h gene knockout mouse model

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