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

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaperJournal articleResearchpeer-review

  • Daniel P Dever, Department of Pediatrics, Stanford University, Stanford, California 94305, USA.
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  • Samantha G Scharenberg, Department of Pediatrics, Stanford University, Stanford, California 94305, USA.
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  • Joab Camarena, Department of Pediatrics, Stanford University, Stanford, California 94305, USA.
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  • Eric J Kildebeck, Department of Pediatrics, Stanford University, Stanford, California 94305, USA.
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  • Joseph T Clark, Department of Pediatrics, Stanford University, Stanford, California 94305, USA.
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  • Renata M Martin, Department of Pediatrics, Stanford University, Stanford, California 94305, USA.
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  • Rasmus O Bak
  • Yuming Tang, StemCells Inc, Newark, CA 94560, USA.
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  • Monika Dohse, StemCells Inc, Newark, CA 94560, USA.
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  • Johannes A Birgmeier, Department of Developmental Biology, Stanford University, Stanford, CA 94305, USA.
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  • Karthik A Jagadeesh, Department of Developmental Biology, Stanford University, Stanford, CA 94305, USA.
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  • 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.
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  • Ann Tsukamoto, ReGen Med Division, BOCO Silicon Valley, Palo Alto, CA 94303, USA.
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  • Natalia Gomez-Ospina, Department of Pediatrics, Stanford University, Stanford, California 94305, USA.
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  • Nobuko Uchida, ReGen Med Division, BOCO Silicon Valley, Palo Alto, CA 94303, USA. Electronic address: nobuko.uchida@bocosiliconvalley.com.
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  • Matthew H Porteus, Department of Pediatrics, Stanford University, Stanford, CA 94305, USA. Electronic address: mporteus@stanford.edu.

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.

Original languageEnglish
JournaliScience
Volume15
Pages (from-to)524-535
Number of pages12
ISSN2589-0042
DOIs
Publication statusPublished - 31 May 2019
Externally publishedYes

    Research areas

  • GALC GENE, HEMATOPOIETIC STEM, LARGE DELETION, NEUROPROTECTION, RNA, TRANSPLANTATION

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