<p style="text-align: justify;">This study investigated pre-service biology teachers' (PSBTs’) technological pedagogical content knowledge (TPACK) development. A TPACK-based technology integration course (TPACK-BTIC) was implemented. The study employed a convergent parallel mixed-methods approach. A TPACK survey questionnaire and interview schedule were used to collect data from 50 PSBTs. The quantitative data were analysed by computing means, standard deviations, and dependent samples t-tests, while qualitative data were analysed using deductive thematic analysis based on the TPACK domains. Findings indicate that the intervention positively affected PSBTs’ TPACK development with significant improvements in technological knowledge (TK), technological pedagogical knowledge (TPK), technological content knowledge (TCK) and overall technological pedagogical content knowledge domains. In contrast, pedagogical content knowledge (PCK) and pedagogical knowledge domains showed no statistically significant improvements. PSBTs’ engagement in microteaching lesson study, reflection on using technology, and collaboratively designing lesson plans improved PSBTs’ TPACK domains. The study recommends that teacher training institutions consider implementing content-based technology integration courses that engage pre-service teachers in microteaching lesson study, reflecting on technology use and collaborative designing of curriculum materials that involve using technology to support their TPACK development.</p>