title: CRISPR/Cas9 Targeted Gene Editing and Cellular Engineering in Fanconi Anemia.
abstract: The ability to rationally target disease-causing mutations has been made possible with programmable nucleases with the CRISPR/Cas9 system representing a facile platform for individualized gene-based medicine. In this study we employed footprint free reprogramming of fibroblasts from a patient with mutations to the Fanconi anemia I (FANCI) gene to generate induced pluripotent stem cells (iPSC). This process was accomplished without gene complementation and the resultant iPSC were able to be gene corrected in a robust manner using the Cas9 nickase. The self-renewing iPSC that were maintained under feeder free conditions were differentiated into cells with characteristics of definitive hematopoiesis. This defined and highly efficient procedure employed small molecule modulation of the hematopoietic differentiation pathway and a vascular induction technique to generate hematopoietic progenitors. In sum, our results demonstrate the ability to induce patient derived FA cells to pluripotency for patient specific therapeutic cell derivation.
authors.affiliation: University of Minnesota Twin Cities, 5635, Pediatrics, Minneapolis, Minnesota, United States.|University of Minnesota Center for Genome Engineering, 189708, Minneapolis, Minnesota, United States.|Stem Cell Institute, Minneapolis, Minnesota, United States ; osbor026@umn.edu.
authors.forename: Mark
authors.initials: M
authors.lastname: Osborn
authors.affiliation: University of Minnesota Twin Cities, 5635, Pediatrics, Minneapolis, Minnesota, United States ; lonet005@umn.edu.
authors.forename: Cara-Lin
authors.initials: CL
authors.lastname: Lonetree
authors.affiliation: University of Minnesota Twin Cities, 5635, Pediatrics, Minneapolis, Minnesota, United States ; webb0178@umn.edu.
authors.forename: Beau R
authors.initials: BR
authors.lastname: Webber
authors.affiliation: University of Minnesota Twin Cities, 5635, Pediatrics, Minneapolis, Minnesota, United States ; ddpatel@umn.edu.
authors.forename: Dharmeshkumar
authors.initials: D
authors.lastname: Patel
authors.affiliation: University of Minnesota Twin Cities, 5635, Pediatrics, Minneapolis, Minnesota, United States ; dunmi002@umn.edu.
authors.forename: Samantha
authors.initials: S
authors.lastname: Dunmire
authors.affiliation: University of Minnesota Twin Cities, 5635, Pediatrics, Minneapolis, Minnesota, United States ; leira001@umn.edu.
authors.forename: Amber N
authors.initials: AN
authors.lastname: McElroy
authors.affiliation: University of Minnesota Twin Cities, 5635, Pediatrics, Minneapolis, Minnesota, United States ; apdefeo@umn.edu.
authors.forename: Anthony P
authors.initials: AP
authors.lastname: DeFeo
authors.affiliation: University of Minnesota Twin Cities, 5635, Pediatrics, Minneapolis, Minnesota, United States ; macmi002@umn.edu.
authors.forename: Margaret L
authors.initials: ML
authors.lastname: MacMillan
authors.affiliation: University of Minnesota Twin Cities, 5635, Pediatrics, Minneapolis, Minnesota, United States ; wagne002@umn.edu.
authors.forename: John
authors.initials: J
authors.lastname: Wagner
authors.affiliation: University of Minnesota Twin Cities, 5635, Pediatrics, Minneapolis, Minnesota, United States ; blaza001@umn.edu.
authors.forename: Bruce R
authors.initials: BR
authors.lastname: Balzar
authors.affiliation: University of Minnesota Twin Cities, 5635, Pediatrics, Minneapolis, Minnesota, United States.|Stem Cell Institute, Minneapolis, Minnesota, United States ; tolar003@umn.edu.
authors.forename: Jakub
authors.initials: J
authors.lastname: Tolar