Biomaterial transfer is used in all biological fields to manipulate cell function and dissect molecular and cellular mechanisms in physiologic and pathologic settings. New, enabling technologies are required to overcome size limitations inherent in all current delivery methods to allow the transfer of large cargo, such as live pathogens, whole chromosomes, and (eventually) bioengineered replacement components, such as genetically modified mitochondria, for new-age therapies. As Nicole Rusk, senior editor Nature Methods, recently opined surprisingly, no methods for efficiently bringing impermeant large molecules into living cells exist (Nat Methods 8:44, 2011). This proposal is focused on our recently developed cargo transfer technology, which is called the photothermal nanoblade, to deliver whole human chromosomes into human pluripotent stem cells (hPSCs) in order to examine chromosome reprogramming. These transferred chromosomes will be analyzed for epigenetic reprogramming within the hPSC nucleus using standard and cutting-edge molecular techniques and is supported by a strong institutional bioinformatics infrastructure. This proposal also aims to develop a microfluidics-integrated photothermal nanoblade platform for next-generation high- throughput and near simultaneous delivery of large cargo into more than 10,000 cells in seconds. Achieving these goals will provide custom engineered hPSCs for biological investigations, an assessment of the chromosome reprogramming capability of federally-registered H1 and H9 human embryonic stem cells (hESCs), and an advanced yet simple to use platform to overcome potential biological obstacles that, if they exist, would limit success due to an insufficient throughput, should chromosome integration be a low frequency event, which is currently not known. PUBLIC HEALTH RELEVANCE: Biomaterial transfer is used in all biological fields to manipulate cell function and dissect molecular and cellular mechanisms in physiologic and pathologic settings. New, enabling technologies are required to overcome size limitations inherent in all current delivery methods to allow the transfer of large cargo for new-age therapies. Our R21 proposal is to use a novel photothermal nanoblade system (Wu, et al., in press, Analytical Chemistry, 2011- see paper in appendix) to deliver whole human chromosomes into pluripotent human stem cells, a currently impossible task, in order to examine whole chromosome epigenetic reprogramming.