Heterozygous and homozygous knock-in mice showed no significant abnormality in any of the above measures at time points up to 2 years. Biochemical, immunological, electron microscopic, fundus autofluorescence, electroretinography and laser photocoagulation analyses were used to characterise the mouse model. We generated and characterised a mouse " knock-in" model carrying the Ser163Arg mutation in the orthologous murine C1qtnf5 gene by site-directed mutagenesis and homologous recombination into mouse embryonic stem cells.
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In this chapter, I describe the generation of epitope-tagged mice using engineered endonuclease and single-stranded oligodeoxynucleotide through the mouse zygote as an example of how to generate a knock-in mouse by genome editing.Ĭharacterisation of a C1qtnf5 Ser163Arg knock-in mouse model of late-onset retinal macular degeneration.ĭirectory of Open Access Journals (Sweden)įull Text Available A single founder mutation resulting in a Ser163Arg substitution in the C1QTNF5 gene product causes autosomal dominant late-onset retinal macular degeneration (L-ORMD in humans, which has clinical and pathological features resembling age-related macular degeneration. Instead of the conventional gene-targeting method using embryonic stem cells, an exogenous DNA sequence can be inserted into the target locus in the zygote using genome editing technology. Knock-in mice are useful for evaluating endogenous gene expressions and functions in vivo. Generation of Knock-in Mouse by Genome Editing. The differentially expressed genes identified in this study may shed new insights into the understanding of the pathogenic mechanism and the phenotypic modification of homozygous p.V37I. Gene expression microarray analysis identified 105 up-regulated and 43 down-regulated genes in P5 knock-in mouse cochleae (P knock-in mouse modeled the hearing phenotype of the human patients and can serve as a useful animal model for further studies. Overall no significant developmental and morphological abnormality was observed in the knock-in mouse cochlea, while confocal immunostaining and electron microscopic scanning revealed minor loss of the outer hair cells. Auditory brainstem response test showed that the knock-in mice developed progressive, mild-to-moderate hearing loss over the first 4-9 months.
To investigate the pathogenic mechanism underlying this variant, we generated a knock-in mouse model of homozygous p.V37I by an embryonic stem cell gene targeting method.
The homozygous p.V37I variant in GJB2 is prevalent in East and Southeast Asians and may lead to mild-to-moderate hearing loss with reduced penetrance. Characterization of a knock-in mouse model of the homozygous p.V37I variant in Gjb2.Ĭhen, Ying Hu, Lingxiang Wang, Xueling Sun, Changling Lin, Xin Li, Lei Mei, Ling Huang, Zhiwu Yang, Tao Wu, Hao
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