Lysine 117 Residue is Essential for the Function of the Hepatocyte nuclear factor 1a

Hepatocyte nuclear factor 1a (HNF1a) play essential roles in controlling development and metabolism, its mutations are clearly linked to the occurrence of maturity-onset diabetes of the young (MODY3) in human. 117 lysine to glutamic acid (E117) mutation in the HNF1a gene was clinically associated with MOYD3, but no functional data on this variant is available. Here, we generated a knock-in mouse model in which the lysine 117 is replaced with glutamic acid in the endogenous murine HNF1a locus. The HNF1aE117/E117 mice reproduced HNF1-null phenotypes including dwarfism, hepatic dysfunction, renal Fanconi syndrome, diabetes and die from a progressive wasting syndrome, but no hyperphenylalaninemia. Moreover, HNF1aE117/E117 mice shared similar hepatic gene expression profiles with HNF1 deficient mice. In vitro experiments showed that HNF1 mutants in which the lysine 117 residue were replaced by glutamic acid, arginine or glutamine residues, respectively, had a significant decreased in overall transactivation and DNA binding capacity. K117 mutants had no effect on its protein stabilities and nuclear localization, but significantly blocked its own association. Our results clearly demonstrate that the lysine 117 residue is essential for the function of the HNF1a, and prove important to identify the molecular basis for diseases associated with defects in HNF1a We then performed gene expression profiling analysis using data obtained from RNA-seq of liver tissue of 3 WT mice and 3 KI mice. Overall design: HNF1a K117E knock-in (HNF1aE117/+E117) mice, harboring the human pathogenic mutation (replacement of K with E) in the mouse ortholog, were generated using the CRISPR/Cas9 system on a C57/BL/6J background. Seven chimeras (HNF1aE117/+) were obtained and then backcrossed with wild-type(WT) C57BL/6J mice. We performed comparative transcriptome analysis by RNAseq of liver tissue from WT and KI mice