Genetic Screening of a New Transgenic Mouse Line Humanized for <i>HLA-A*02:01:01:01</i> and <i>hβ2m</i>

Abstract

The development of new humanized transgenic mouse biomodels with the HLA-A*02:01:01:01 gene requires effective methods for target transgene verification in the animal genome. In the present study, we develop a system for genetic screening of animals based on real-time PCR and using highly specific primers to detect all functionally significant parts of the genetic construct. In addition, the Sanger sequencing method showed the absence of chimerism and complete correspondence between the primary nucleotide sequence of the HLA A*02:01:01:01 transgene and the developed engineered genetic construct and human gene HLA A*02:01:01:01. Based on the results of selection and genetic works with the resulting transgenic animals, three most promising sublines were identified. These lines are currently used for breeding a new line of humanized transgenic mice with the HLA-A*02:01:01:01 gene.