Abstract
Corticotropin releasing hormone (CRH) is known to be a major regulator of the HPA axis, but its role in relation to other modulators of ACTH secretion, i.e. vasopressin and catecholamines, is unclear. CRH has also been found in the cerebral cortex and immune system, where its functions are poorly understood. To address these questions, and also to analyze the role of CRH in development of the HPA axis we are constructing a mammalian model of CRH deficiency. We have cloned and sequenced the mouse CRH gene, and mapped its location in the mouse genome to the proximal region of chromosome 3 by interspecific mouse backcrosses. No known mutations near this region have a phenotype suggestive of CRH deficiency. We have constructed a vector which replaces the entire coding region of the CRH gene with the neoR gene and has the herpes thymidine kinase gene flanking CRH sequences. This vector was linearized and electroporated into 6×107 ES cells, which subsequently underwent selection with G418 and gancyclovir. 250 doubly resistant clones were obtained, and 2 targeted mutations with disrupted CRH genes were obtained from 87 screened. These targeted clones will be injected into blastocysts to obtain chimeric animals, which will then be bred to obtain animals hetero- and homozygous for CRH deficiency.
Article PDF
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Muglia, L., Copeland, N., Jenkins, N. et al. TARGETED MUTATION OF THE MOUSE CRH GENE IN EMBRYONIC STEM (ES) CELLS. Pediatr Res 33 (Suppl 5), S31 (1993). https://doi.org/10.1203/00006450-199305001-00166
Issue Date:
DOI: https://doi.org/10.1203/00006450-199305001-00166