Rapid and precise genotyping of transgene zygosity in mice using an allele-specific method.

ABSTRACT: Precise determination of transgene zygosity is essential for use of transgenic mice in research. Because integration loci of transgenes are usually unknown due to their random insertion, assessment of transgene zygosity remains a challenge. Current zygosity genotyping methods (progeny testing, qPCR, and NGS-computational biology analysis) are time consuming, prone to error or technically challenging. Here, we developed a novel method to determine transgene zygosity requiring no knowledge of transgene insertion loci. This method applies allele-specific restriction enzyme digestion of PCR products (RE/PCR) to rapidly and reliably quantify transgene zygosity. We demonstrate the applicability of this method to three transgenic strains of mice (Atm Tg^C3001L, Nes-Cre, an

SUBMITTER: Yang J 

PROVIDER: S-EPMC10087101 | biostudies-literature | 2023 Jun

REPOSITORIES: biostudies-literature