EAGER: Biomanufacturing: CRISPR to increase the homogeneity and efficiency of stem cell differentiation

Michigan State University, East Lansing MI

Investigators

Abstract

PI: Chan, Christina Proposal Number: 1547518 Stem cells are promising for the regeneration of aged, injured and diseased tissues and organs, but to realize their therapeutic potential growth of unwanted cell types should be arrested, and improvement in efficiency of converting them to a specific cell type is necessary. The proposed work will examine a genome editing approach for eliminating growth of unwanted cell lineages. The overall goal of this project is to increase the efficiency and specificity of stem cell differentiation by reducing off-target cell types through CRISPR. CRISPR/Cas9 mediated knockout of transcription factors involved in the maturation of serotonergic, glutamatergic and GABAergic phenotypes. Clones will be DNA sequenced to confirm deletions in the regions involved in serotonin, glutamate and GABA synthesis pathways, and those lacking these genes will be treated with FI (forskolin and IBMX) to induce a higher percentage of cells to express a dopaminergic phenotype. This method will also be applied to increase the efficiency and specificity of neural stem cell differentiation. The results are anticipated to easily transfer to induced pluripotent, embryonic, and adult stem cells. From an educational standpoint, this project will provide interdisciplinary training to students with an opportunity to integrate knowledge from different fields to investigate non-traditional engineering problems. The investigators will recruit and retain female and minority engineering students into the research project.