University teacher education aims at developing sustained knowledge and positive attitudes towards evidence-based instructional concepts. This experimental study examined (a) if two e-learning formats help develop sustained knowledge and positive attitudes towards evidence-based reading instruction, (b) if self-regulative principles can boost these effects, and (c) if effects are moderated by learner characteristics (namely prior knowledge, self-efficacy, learning preferences). N = 100 student teachers participated in one of two different e-learning formats on evidence-based reading instruction. While one group attended an e-learning course with self-regulative elements (adaptivity and feedback), the other group watched an e-lecture without these self-regulative elements. In both groups, participants showed positive attitudes and mastery of declarative knowledge, as the majority reached a minimum of 80% in criterion-oriented tests. However, participants’ procedural knowledge was less pronounced. A MANOVA showed no significant group differences between the e-learning course with self-regulative elements and the e-lecture without self-regulative elements with regard to knowledge and attitude acquisition. Path analyses displayed that individual learner characteristics (prior knowledge, self-efficacy, learning preferences) had an impact on attitudes, but not on knowledge acquisition. Implications for university education are discussed in the context of combining e-learning formats with face-to-face courses to also achieve mastery of procedural knowledge and to strengthen the effect of self-regulative elements.