CRISPR/Cas9 Genome Engineering in Engraftable Human Brain-Derived Neural Stem Cells

Daniel P. Dever; Samantha G. Scharenberg; Joab Camarena; Eric J. Kildebeck; Joseph T. Clark; Renata M. Martin; Rasmus O. Bak; Yuming Tang; Monika Dohse; Johannes A. Birgmeier; Karthik A. Jagadeesh; Gill Bejerano; Ann Tsukamoto; Natalia Gomez-Ospina; Nobuko Uchida; Matthew H. Porteus

iScience (May 2019)

CRISPR/Cas9 Genome Engineering in Engraftable Human Brain-Derived Neural Stem Cells

Daniel P. Dever,
Samantha G. Scharenberg,
Joab Camarena,
Eric J. Kildebeck,
Joseph T. Clark,
Renata M. Martin,
Rasmus O. Bak,
Yuming Tang,
Monika Dohse,
Johannes A. Birgmeier,
Karthik A. Jagadeesh,
Gill Bejerano,
Ann Tsukamoto,
Natalia Gomez-Ospina,
Nobuko Uchida,
Matthew H. Porteus

Affiliations

Daniel P. Dever: Department of Pediatrics, Stanford University, Stanford, CA 94305, USA
Samantha G. Scharenberg: Department of Pediatrics, Stanford University, Stanford, CA 94305, USA
Joab Camarena: Department of Pediatrics, Stanford University, Stanford, CA 94305, USA
Eric J. Kildebeck: Department of Pediatrics, Stanford University, Stanford, CA 94305, USA
Joseph T. Clark: Department of Pediatrics, Stanford University, Stanford, CA 94305, USA
Renata M. Martin: Department of Pediatrics, Stanford University, Stanford, CA 94305, USA
Rasmus O. Bak: Department of Pediatrics, Stanford University, Stanford, CA 94305, USA
Yuming Tang: StemCells Inc, Newark, CA 94560, USA
Monika Dohse: StemCells Inc, Newark, CA 94560, USA
Johannes A. Birgmeier: Department of Developmental Biology, Stanford University, Stanford, CA 94305, USA
Karthik A. Jagadeesh: Department of Developmental Biology, Stanford University, Stanford, CA 94305, USA
Gill Bejerano: Department of Pediatrics, Stanford University, Stanford, CA 94305, USA; Department of Developmental Biology, Stanford University, Stanford, CA 94305, USA; Department of Computer Science, Stanford University, Stanford, CA 94305, USA
Ann Tsukamoto: ReGen Med Division, BOCO Silicon Valley, Palo Alto, CA 94303, USA
Natalia Gomez-Ospina: Department of Pediatrics, Stanford University, Stanford, CA 94305, USA
Nobuko Uchida: ReGen Med Division, BOCO Silicon Valley, Palo Alto, CA 94303, USA; Corresponding author
Matthew H. Porteus: Department of Pediatrics, Stanford University, Stanford, CA 94305, USA; Corresponding author

Journal volume & issue: Vol. 15
pp. 524 – 535

Abstract

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Summary: Human neural stem cells (NSCs) offer therapeutic potential for neurodegenerative diseases, such as inherited monogenic nervous system disorders, and neural injuries. Gene editing in NSCs (GE-NSCs) could enhance their therapeutic potential. We show that NSCs are amenable to gene targeting at multiple loci using Cas9 mRNA with synthetic chemically modified guide RNAs along with DNA donor templates. Transplantation of GE-NSC into oligodendrocyte mutant shiverer-immunodeficient mice showed that GE-NSCs migrate and differentiate into astrocytes, neurons, and myelin-producing oligodendrocytes, highlighting the fact that GE-NSCs retain their NSC characteristics of self-renewal and site-specific global migration and differentiation. To show the therapeutic potential of GE-NSCs, we generated GALC lysosomal enzyme overexpressing GE-NSCs that are able to cross-correct GALC enzyme activity through the mannose-6-phosphate receptor pathway. These GE-NSCs have the potential to be an investigational cell and gene therapy for a range of neurodegenerative disorders and injuries of the central nervous system, including lysosomal storage disorders. : Molecular Biology; Neuroscience; Bioengineering; Biotechnology; Cell Biology; Biological Sciences Research Methodologies Subject Areas: Molecular Biology, Neuroscience, Bioengineering, Biotechnology, Cell Biology, Biological Sciences Research Methodologies

Published in iScience

ISSN: 2589-0042 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science
Website: http://www.cell.com/iscience/home

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