A novel carcinogenic mouse model by site-directed insertion of tandem human HRAS large DNA fragment into 15E1 site.

ABSTRACT:

Background

The precise insertion of large DNA fragments (>3-5 kb) remains one of the key obstacles in establishment of genetically modified murine models.

Methods

A 21 kb large DNA fragment containing three tandemly linked copies of the human HRAS gene was inserted into the genome of C57BL/6J mouse, generating a mouse model designated as KI.C57-ras (or named NF-hHRAS). Whole-genome sequencing and Sanger sequencing were utilized to it confirm precise insertion and copy number. The stability of transgene expression among different generations was verified from multiple aspects using by digital PCR, western blot and DNA sequencing. To assess tumor susceptibility in the mouse model, N-Nitroso-N-methylurea (MNU) was administered at a dosage of 75 mg/kg. Histopathological examinat

SUBMITTER: Liu S 

PROVIDER: S-EPMC12746206 | biostudies-literature | 2025 Nov

REPOSITORIES: biostudies-literature