title: RNA-Generated and Gene-Edited Induced Pluripotent Stem Cells for Disease Modeling and Therapy.
abstract: Cellular reprogramming by epigenomic remodeling of chromatin holds great promise in the field of human regenerative medicine. As an example, human-induced Pluripotent Stem Cells (iPSCs) obtained by reprograming of patient somatic cells are sufficiently similar to embryonic stem cells (ESCs) and can generate all cell types of the human body. Clinical use of iPSCs is dependent on methods that do not utilize genome altering transgenic technologies that are potentially unsafe and ethically unacceptable. Transient delivery of exogenous RNA into cells provides a safer reprogramming system to transgenic approaches that rely on exogenous DNA or viral vectors. RNA reprogramming may prove to be more suitable for clinical applications and provide stable starting cell lines for gene-editing, isolation, and characterization of patient iPSC lines. The introduction and rapid evolution of CRISPR/Cas9 gene-editing systems has provided a readily accessible research tool to perform functional human genetic experiments. Similar to RNA reprogramming, transient delivery of mRNA encoding Cas9 in combination with guide RNA sequences to target specific points in the genome eliminates the risk of potential integration of Cas9 plasmid constructs. We present optimized RNA-based laboratory procedure for making and editing iPSCs. In the near-term these two powerful technologies are being harnessed to dissect mechanisms of human development and disease in vitro, supporting both basic, and translational research. J. Cell. Physiol. 232: 1262-1269, 2017. © 2016 Wiley Periodicals, Inc.
authors.affiliation: ITB-CNR, Segrate, Milan, Italy.|National Institutes of Health, NIDDK, Laboratory of Cell and Molecular Biology, Rockville Pike, Bethesda, Maryland.|MTI-GlobalStem, Gaithersburg, Maryland.
authors.forename: James
authors.initials: J
authors.lastname: Kehler
authors.affiliation: ITB-CNR, Segrate, Milan, Italy.
authors.forename: Marianna
authors.initials: M
authors.lastname: Greco
authors.affiliation: ITB-CNR, Segrate, Milan, Italy.
authors.forename: Valentina
authors.initials: V
authors.lastname: Martino
authors.affiliation: MTI-GlobalStem, Gaithersburg, Maryland.
authors.forename: Manickam
authors.initials: M
authors.lastname: Pachiappan
authors.affiliation: MTI-GlobalStem, Gaithersburg, Maryland.
authors.forename: Hiroko
authors.initials: H
authors.lastname: Yokoe
authors.affiliation: Organovo, San Diego, California.
authors.forename: Alice
authors.initials: A
authors.lastname: Chen
authors.affiliation: Ascendance Biotechnologies, Medford, Maryland.
authors.forename: Miranda
authors.initials: M
authors.lastname: Yang
authors.affiliation: MTI-GlobalStem, Gaithersburg, Maryland.
authors.forename: Jonathan
authors.initials: J
authors.lastname: Auerbach
authors.affiliation: MTI-GlobalStem, Gaithersburg, Maryland.
authors.forename: Joel
authors.initials: J
authors.lastname: Jessee
authors.affiliation: Department of Gynecology and Obstetrics, Muenster University Hospital, Muenster, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Gotte
authors.affiliation: ITB-CNR, Segrate, Milan, Italy.
authors.forename: Luciano
authors.initials: L
authors.lastname: Milanesi
authors.affiliation: ITB-CNR, Segrate, Milan, Italy.
authors.forename: Alberto
authors.initials: A
authors.lastname: Albertini
authors.affiliation: Department of Biology, College of Science and Technology, Temple University, Philadelphia, Pennsylvania.|Sbarro Institute for Cancer Research and Molecular Medicine, College of Science and Technology, Temple University, Philadelphia, Pennsylvania.
authors.forename: Gianfranco
authors.initials: G
authors.lastname: Bellipanni
authors.affiliation: ITB-CNR, Segrate, Milan, Italy.
authors.forename: Ileana
authors.initials: I
authors.lastname: Zucchi
authors.affiliation: ITB-CNR, Segrate, Milan, Italy.
authors.forename: Rolland A
authors.initials: RA
authors.lastname: Reinbold
authors.affiliation: Department of Biology, College of Science and Technology, Temple University, Philadelphia, Pennsylvania.|Sbarro Institute for Cancer Research and Molecular Medicine, College of Science and Technology, Temple University, Philadelphia, Pennsylvania.
authors.forename: Antonio
authors.initials: A
authors.lastname: Giordano
mesh_terms: D002454:Cell Differentiation; D065150:Cellular Reprogramming; D004194:Disease; D055808:Drug Discovery; D000072669:Gene Editing; D005822:Genetic Vectors / Q000378:metabolism; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000378:metabolism*; D008954:Models, Biological; D057285:Precision Medicine; D012313:RNA / Q000378:metabolism*
chemical_list: D012313:RNA