<p>In today’s educational environments, videos have become integral to learning and teaching processes. The rapid spread of videos in education necessitates the development of video course designs that consider individual differences and learner needs. However, generic designs fail to address the varied needs of individuals. Video lectures that consider individual differences can help students benefit more effectively from the materials. However, it is observed that the literature on how learners with different cognitive characteristics benefit from video designs and how individual learning processes can be supported is limited, and research in this field is still at an early stage. Therefore, this research investigates the effects of pre-service science teachers’ sustained attention levels and different video lecture types on their learning and recall performance on the cell cycle. The research was conducted with a 2 × 2 between-group factorial design, which is one of the experimental methods. The research participants included 100 pre-service science teachers. During the data collection process, a computer-based sustained attention test, an achievement test, and the GazePoint GP3 eye tracking system were used. Two-way ANOVA and the Mann–Whitney U test were used for data analysis. Results show that pre-service science teachers’ learning and recall performance differ significantly depending on the type of interaction in video lectures and their sustained attention levels. In addition, participants with high sustained attention levels achieved the highest learning performance when they watched picture-in-picture video lectures. On the other hand, the lowest learning performance was observed in the group where participants with low sustained attention levels watched lecture videos with voiceover. The results from eye tracking data also support this finding.</p>