期刊
GENESIS
卷 60, 期 6-7, 页码 -出版社
WILEY
DOI: 10.1002/dvg.23479
关键词
ARX; brain development; CRSPR; cas9; detection of FLAG tagged-endogenous protein; Flag-Arx knock-in mouse
资金
- National Center of Neurology and Psychiatry
- National Institute of Neurological Disorders and Stroke [R01NS100007]
The ARX gene is crucial for brain development, and mutations can lead to neurodevelopmental disorders. Mouse models have provided insights into the pathophysiology of these disorders. The FLAG-Arx mouse line is a valuable tool for isolating ARX complexes.
The Aristaless-related homeobox (ARX) is a paired-like homeodomain transcription factor playing important roles in brain development. Patients with mutations in ARX have a spectrum of neurodevelopmental disorders such as epilepsy, intellectual disability, and autism spectrum disorder, with or without structural abnormalities of the brain such as lissencephaly (smooth brain), microcephaly (small brain), and/or agenesis of the corpus callosum. Mouse models have provided important clues on the pathophysiologic roles of ARX in these disorders. However, successfully isolating specific in vivo complexes of ARX, with DNA and proteins, has remained as a challenge. To facilitate in vivo detection of ARX complexes, we generated a mouse line containing one epitope of FLAG-tag (1 x FLAG) targeted at the translational start site of the endogenous Arx gene using CRSPR/Cas9 strategy. Homozygous Flag-Arx mice are viable and fertile without gross abnormality, suggesting that the FLAG-tag does not perturb the normal function of ARX. Using a FLAG antibody, we successfully detected ARX with immunofluorescent staining and pulled down ARX in embryonic brain tissues. This Flag-Arx mouse line will be a useful tool to isolate ARX complexes from mouse tissues for many applications.
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