Engineering non-viral gene delivery methods for iPSC-derived Alzheimer’s models

Induced pluripotent stem cells (iPSCs) have emerged as a powerful tool for regenerative medicine and disease modelling, particularly for complex, age-related disorders like Alzheimer’s disease. However, reprogramming efficiencies remain extremely low, and iPSC’s often exhibit high heterogeneity in genomic stability and epigenetic status. These factors critically impact downstream differentiation quality and reproducibility. Additionally, studies have demonstrated that reprogramming is highly stressful for cells, causing DNA damage, and that reprogramming machineries are intertwined with mechanisms controlling DNA damage response, apoptosis and cell cycle. For example, the process of iPSC generation and reprogramming has been demonstrated to be facilitated by cell-death machinery such as the apoptosis-inducing caspases. Further, downregulation of TP53, the ‘cellular gatekeeper’ has also been shown to increase reprogramming efficiencies, although complete knockout has been associated with genomic instability. Although non-integrative vectors such as Sendai virus or episomal plasmids aim to reduce mutational burden, whole-genome sequencing reveals that reprogramming still introduces genetic aberrations. Fundamentally, the molecular mechanisms underpinning cellular reprogramming remain poorly understood and there is little to no information regarding how the delivery method and agent itself impacts the intracellular responses such as DNA damage, immune activation, and cell fate decisions.

This project aims to engineer precise, non-viral gene delivery systems using a technique called ‘DNA origami' to address these challenges and advance iPSC technologies for Alzheimer’s disease modelling. The student will develop a library of DNA origami constructs encoding transcription factors and epigenetic editing tools. Using cutting-edge approaches including super-resolution imaging, NanoSIMS, and single-cell multi-omics (including RNA-seq), the intracellular uptake, trafficking, and cellular responses to DNA origami will be characterised. In particular, the intracellular protein corona will be investigated by delivering DNA origami, fixing cells at various time points, performing pulldown assays, and analysing bound proteins by LC-MS/MS. Innate immune activation, such as through the cGAS-STING pathway, and DNA damage responses will be assessed through imaging and molecular assays. Dual DNA barcodes encoded in the origami constructs will enable multiplexed single-cell tracking of both delivery and gene expression. This comprehensive, correlative approach will provide critical insights into the intracellular barriers of non-viral delivery, enabling more efficient, stable, and scalable iPSC reprogramming for Alzheimer’s disease research.

This scholarship is part of the MIND-AD CRE Scholarship program which will support three students across its advertised projects. Applicants will be required to submit an online application and attend an interview.