Additional mutation search
To investigate the presence of additional mutations that
could be associated with PD or other inherited diseases,
we analyzed genomic DNA from each of the fibroblast lines
using the Ion AmpliSeq™ Inherited Disease Panel on the
Ion PGM™ Sequencer. This panel provides highly multiplexed
target selection of exons of 325 genes that are mutated in
over 700 inherited diseases, according to the NCBI ClinVar
database [6], and comprise genes implicated in some of the
most common forms of inherited deafness, blindness, heart
disease, Parkinson's disease, immunodeficiency, various
ataxias, anemias, and treatable metabolic syndromes. PDrelated genes in the panel include LRRK2, FBX07, PINK1,
MAPT, SNCA, and TAF1.
Briefly, targeted libraries were generated from genomic
DNA isolated from donor fibroblasts, then transferred to
the Ion OneTouch™ System for template preparation, then
sequenced on the Ion PGM™ Sequencer. Automated analysis
was performed with Torrent Suite™ Software on the Torrent
Server. Our results with the Ion AmpliSeq™ Inherited Disease
Panel confirmed the presence of the previously detected
LRRK2 and/or GBA mutation in the expected lines, and no
unexpected mutations in other lines, including the MSA line
that had been previously genotyped with a broader panel
of PD-related genes and in which no PD-related mutations
were identified.
10 Life Technologies | Parkinson's cell model
Instrument highlight—Ion PGM™ Sequencer
The Ion PGM™ Sequencer was used to evaluate the
donor fibroblasts for additional mutations prior to
reprogramming. The fibroblasts were found to have
the previously detected LRRK2 and/or GBA mutation
in the expected lines, and no unexpected mutations.
