A Novel Approach to Purifying iPSC Derived Cell Cultures

DESCRIPTION provided by applicant We are pioneering an RNA based switch technology called structurally interacting RNAs sxRNA which utilizes post transcriptional gene regulation as a reporter for miRNA detection Normally RNA binding proteins RBP associate with a andapos stem loop structure to facilitate translation of an upstream coding region by as much as an order of magnitude It is possible to modify the mRNA to modulate translation of the reporter gene by controlling the binding of RBP This is accomplished by creating a trans acting three way junction with an additional RNA such as a miRNA binds and stabilizes the functional structure by base pairing with the flanking regions of the custom designed stem loop Our goal with this proposal is use this selective expression capability to create homogenous populations of cells in culture focusing specifically on stem cells We plan to create a procedure sold as a kit that would eliminate unwanted cells from a differentiated cell population We will accomplish this by using miRNA expression in the desired cells to turn on translation of an antibiotic gene thereby maintaining the desired cells Success with this product development effort will result in a product that has broad appeal to stem cell researchers both for the benefits of increased homogeneity of stem cell cultures and potentially faster development of stem cell related lines and position us to further enhance the technology for use in CHO cells lines for cGMP cell culturing PUBLIC HEALTH RELEVANCE HocusLocus is developing a novel technology to control the expression of an mRNA This technology can be used express an antibiotic resistance gene only in the presence of cells that express a particular miRNA The product being developed will allow stem cell researchers to ensure the homogeneity and potentially speed up culturing of stem cells or differentiated cells by killing unwanted cells based on their miRNA expression patterns