Generation of isogenic pluripotent stem cells differing exclusively at two early onset Parkinson point mutations

By F. Soldner, J. Laganière, A. Cheng, D. Hockemeyer, Qing Gao, R. Alagappan, V. Khurana, L. Golbe, R. Myers, Susan Lindquist and 10 more. Nature Reviews Neuroscience, 2011

754 citations recorded by the source corpus.

SUMMARY Patient-specific induced pluripotent stem cells (iPSCs) derived from somatic cells provide a unique tool for the study of human disease, as well as a promising source for cell-replacement therapies. However one crucial limitation has been the inability to perform experiments under genetically defined conditions. This is particularly relevant for late age-onset disorders where in vitro phenotypes are predicted to be subtle and susceptible to significant effects of genetic background variations. By combining zinc-finger nuclease (ZFN)-mediated genome editing and iPSC technology we provide a generally applicable solution to this problem by generating sets of isogenic disease and control human pluripotent stem cells that differ exclusively at either of two susceptibility variants for Parkinson’s disease by modifying the underlying point mutations (A53T/E46K) in the α-synuclein gene. The robust capability to genetically correct disease-causing point mutations in patient-derived hiPSCs represents not only a significant progress for basic biomedical research but also a major advancement towards hiPSC-based cell-replacement therapies.

Subfields: Cellular and Molecular Neuroscience, Developmental Neuroscience, Molecular Biology

View Paper: https://www.semanticscholar.org/paper/c33e6ee38b5c884c2b34795e0687d73b83c575d4

How researchers see this paper

The researchers whose trusted networks rank it highest, strongest advocate first. Every researcher gets a different order.

In Cellular and Molecular Neuroscience

In Molecular Biology

Citations in the corpus

Double nicking by RNA-guided CRISPR Cas9 for enhanced genome editing specificity (2013, background): “These unwantedmutations can potentially limit the utility of Cas9 for genome editing applications that require high levels of precision, such as generation of isogenic cell lines for testing causal genetic variations (Soldner et al., 2011) or in vivo and ex vivo genome-editing-based therapies.”
Microglial cell origin and phenotypes in health and disease (2011, background): “Interest in the potential utility of studying patient-derived microglia has been raised by the recently accrued capacity to model neurodegenerative diseases using patient-derived induced pluripotent stem (iPS) cells that can give rise to neurons and astrocytes with disease phenotypes in cultur”
Beyond GWASs: illuminating the dark road from association to function. (2013, methodology): “used engineered ZFNs to generate isogenic pluripotent stem cells to study risk variants for Parkinson disease (MIM 168600).(107) The lines differed by two single point mutations in a-synuclein (MIM 163890) but maintained a pluripotent state, the ability to form teratomas, and the ability to differen”
Delivering CRISPR: a review of the challenges and approaches (2018, background): “Both gene correction (Figure 2(C)) and gene addition (Figure 2(D)) require an exogenous DNA template that can be introduced as either single-stranded (Radecke et al., 2010; Chen et al., 2011; Soldner et al., 2011) or double-stranded DNA (Rouet et al.”