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November 07-08, 2019 | Melbourne, Australia

Molecular Biology and Genetic Engineering

International Conference on

Journal of Research and Reports on Genetics | Volume 3

Mitochondrial DNA structural variation sequencing from single cell (MitoSV-seq)

Elham Jaberi, Emilie Tresse, Joachim Weischenfeldt

and

Shohreh Issazadeh-Navikas

University of Copenhagen, Denmark

itochondrialDNA(mtDNA)pointmutationsandstructural

variations (SVs) have been reported to contribute to

axonal and neuronal cell death and therefore diseases of the

central nervous system. Both technical limitations and the

heteroplasmic nature of mtDNA have made the identification

of SVs in mtDNA challenging. We have developed a novel,

high-resolution method, hereby called MitoSV-seq, to identify

mtDNA-SVs and single nucleotide variations (SNVs) from single

cells in this case in neurons by next generation sequencing,

using a designed positive control. We isolated single

neurons from brain of Ifnb–/–, a mouse developing a sever

demyelinating experimental autoimmune encephalomyelitis

and spontaneous neurodegenerationusing Flowcytometry cell

sorting. mtDNA was amplified exponentially at multiple sites

using rolling circle amplification before sequencing. We found

15 SNVs and 39 SVs exclusively in 70% of Ifnb–/– neurons.

Compared with previous methods, MitoSV-seq is optimized to

identify a higher proportionwith better resolution of variations

with low heteroplasmy in single cells. This could serve as an

efficient way to identify mtDNA-SVs in a range of conditions

that primarily affect mitochondrial functions in the brain.

Speaker Biography

Elham Jaberi has completed her Ph.D. in Cell and molecular biology.

During her PhD thesis, she investigated the genetic and molecular

basis of two neurodegenerative diseases, Parkinson’s disease and

ataxia. During her first postdoc experience, she found two novel genes

associated with mental retardation, and Parkinson’s disease. During her

second postdoc at the BRIC, University of Copenhagen, she is working

on a new animal model for Parkinson’s disease like dementia; IFN-ß

knockout mice (Ifnb–/–). She has 9 published and two submitted papers.

Elham Jaberi et al.

, J Res Rep Genet 2019, Volume 3

DOI: 10.35841/2591-7986-C1-002