Cancer stem cells (CSCs) are a subpopulation of malignant cells with self-renewing capacity, hence being vital in the areas of tumour initiation and progression, therapeutic resistance, and tumour relapse following treatment. Here, we highlight the key transcription factors involved in the production of these CSCs, namely, SOX2, OCT4, Nanog, KLF4 and c-Myc. All of these factors are involved in somatic cell reprogramming, hence inducing pluripotency and leading to the development of many cancers. Due to their roles in the propagation of many cancers, these transcription factors have been implicated as important diagnostic and prognostic tools for cancer patients, enabling a bigger picture understanding of a patient’s cancer presentation. These factors also play pivotal roles in chemoresistance, proving a massive barrier to modern cancer therapies and increasing the rates of cancer recurrence following remission.