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Development and characterization of an Otp conditional loss of function allele
Author(s) -
Hu Yu,
Li Jiamin,
Zhu Yuyuan,
Li Mengqi,
Lin Jianbang,
Yang Lixin,
Wang Chuan,
Lu Zhonghua
Publication year - 2020
Publication title -
genesis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.093
H-Index - 110
eISSN - 1526-968X
pISSN - 1526-954X
DOI - 10.1002/dvg.23370
Subject(s) - conditional gene knockout , biology , homeobox , microbiology and biotechnology , crispr , knockout mouse , loss function , allele , exon , embryonic stem cell , cell type , gene knockout , gene knockin , transcription factor , gene , phenotype , genetics , cell
Summary Orthopedia (Otp) is a homeodomain transcription factor that plays an essential role in the development of hypothalamic neurosecretory systems. Loss of Otp results in the failure of differentiation of key hypothalamic neuroendocrine cell types, and pups die soon after birth. Although the constitutive knockout Otp mouse model ( Otp KO ) has significantly expanded our understanding of Otp's function in vivo, a conditional loss of function Otp allele that enables tissue or cell‐type specific ablation of Otp has not been developed. Here, we used CRISPR/Cas9 gene‐editing technology to generate a conditional Otp knockout mouse line in which exon 2 of the murine Otp gene is flanked by LoxP sites ( Otp f/f ). Crossing the Otp f/f mouse with Agrp‐Ires‐cre mouse, we demonstrate the requirement for Otp in the continuous differentiation of AgRP neurons after cell fate determination. We also show that the residual AgRP neurons in Agrp‐Ires‐cre;Otp f/f mice project to similar downstream target regions. This newly developed Otp f/f mouse can be used to explore the functions of Otp with cell‐type or temporal specificity.