Generation of RAB39B knockout isogenic human embryonic stem cell lines to model RAB39B-mediated Parkinson's disease

Yujing Gao; Gabrielle R Wilson; Kiymet Bozaoglu; Andrew G Elefanty; Edouard G Stanley; Mirella Dottori; Paul J Lockhart

doi:10.1016/j.scr.2018.02.015

Generation of RAB39B knockout isogenic human embryonic stem cell lines to model RAB39B-mediated Parkinson's disease

Stem Cell Res. 2018 Apr:28:161-164. doi: 10.1016/j.scr.2018.02.015. Epub 2018 Feb 21.

Authors

Yujing Gao¹, Gabrielle R Wilson¹, Kiymet Bozaoglu¹, Andrew G Elefanty², Edouard G Stanley³, Mirella Dottori⁴, Paul J Lockhart⁵

Affiliations

¹ Bruce Lefroy Centre for Genetic Health Research, Murdoch Children's Research Institute, Melbourne, Victoria, 3052, Australia; Department of Paediatrics, The University of Melbourne, Melbourne, Victoria, 3052, Australia.
² Department of Paediatrics, The University of Melbourne, Melbourne, Victoria, 3052, Australia; Blood Cell Development and Disease Laboratory, Murdoch Children's Research Institute, The Royal Children's Hospital, Melbourne, Victoria, 3052, Australia; Department of Anatomy and Developmental Biology, Faculty of Medicine, Nursing and Health Sciences, Monash University, Clayton, Victoria, Australia.
³ Department of Paediatrics, The University of Melbourne, Melbourne, Victoria, 3052, Australia; Stem Cell Technology Laboratory, Murdoch Children's Research Institute, The Royal Children's Hospital, , Melbourne, Victoria, 3052, Australia; Department of Anatomy and Developmental Biology, Faculty of Medicine, Nursing and Health Sciences, Monash University, Clayton, Victoria, Australia.
⁴ Centre for Neural Engineering, The University of Melbourne, Carlton, VIC, 3010, Australia; Illawarra Health and Medical Research Institute, Centre for Molecular and Medical Bioscience, University of Wollongong, Wollongong, New South Wales, 2500, Australia.
⁵ Bruce Lefroy Centre for Genetic Health Research, Murdoch Children's Research Institute, Melbourne, Victoria, 3052, Australia; Department of Paediatrics, The University of Melbourne, Melbourne, Victoria, 3052, Australia. Electronic address: [email protected].

PMID: 29499499
DOI: 10.1016/j.scr.2018.02.015

Abstract

Mutations in RAB39B are a known cause of X-linked early onset Parkinson's disease. Isogenic human embryonic stem cell lines carrying two independent deletions of RAB39B were generated using CRISPR/Cas9 genome editing tool. The deletions were confirmed by PCR and direct sequence analysis in two edited stem cell lines. Both cell lines showed pluripotency and displayed a normal karyotype. Further, they were able to form embryoid bodies in vitro, and express markers indicative of differentiation to the three germ layers.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Culture Techniques / methods*
Cell Line
Embryoid Bodies / metabolism
Gene Knockout Techniques*
Human Embryonic Stem Cells / cytology*
Humans
Karyotyping
Mice
Mycoplasma / isolation & purification
Parkinson Disease / genetics*
Parkinson Disease / pathology*
rab GTP-Binding Proteins / genetics*

Substances

Rab39B protein, human
rab GTP-Binding Proteins