Year: 2025
Type: article
Abstract: Precision cancer modeling relies on high-fidelity PCR technology for validating genetically engineered animal models. However, suboptimal primer design frequently causes non-specific amplification and reduced detection sensitivity. In this study, we employed traditional primer design principles to develop target-specific primers for validating Smad4 and Cre gene knockout in a colorectal cancer mouse model. Through optimized PCR amplification and DNA gel electrophoresis, we achieved high specificity in genotyping with clear band separation and confirmed tissue-specific recombinase activity. These results demonstrate that rational primer design ensures reliable amplification efficiency and minimizes off-target effects. Furthermore, we discuss the limitations of conventional design strategies in complex genomic contexts and propose computational approaches for enhancing primer specificity in future studies.
Source: Transactions on Cancer
Authors: Chenyu Bi, Tianyue Liu
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: diamond
OpenAlex ID: W4414475146
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