Generation of SNCA Cell Models Using Zinc Finger Nuclease (ZFN) Technology for Efficient High-Throughput Drug Screening.

PloS One
Warunee DansithongDuong P Huynh

Abstract

Parkinson's disease (PD) is a progressive neurodegenerative disorder caused by loss of dopaminergic neurons of the substantia nigra. The hallmark of PD is the appearance of neuronal protein aggregations known as Lewy bodies and Lewy neurites, of which α-synuclein forms a major component. Familial PD is rare and is associated with missense mutations of the SNCA gene or increases in gene copy number resulting in SNCA overexpression. This suggests that lowering SNCA expression could be therapeutic for PD. Supporting this hypothesis, SNCA reduction was neuroprotective in cell line and rodent PD models. We developed novel cell lines expressing SNCA fused to the reporter genes luciferase (luc) or GFP with the objective to enable high-throughput compound screening (HTS) for small molecules that can lower SNCA expression. Because SNCA expression is likely regulated by far-upstream elements (including the NACP-REP1 located at 8852 bp upstream of the transcription site), we employed zinc finger nuclease (ZFN) genome editing to insert reporter genes in-frame downstream of the SNCA gene in order to retain native SNCA expression control. This ensured full retention of known and unknown up- and downstream genetic elements controlling SNCA ex...Continue Reading

References

Dec 28, 1999·Proceedings of the National Academy of Sciences of the United States of America·K M CrowderS M Bajjalieh
Jun 6, 2000·Journal of Biomolecular Screening·J H ZhangK R Oldenburg
Jul 18, 2002·Proceedings of the National Academy of Sciences of the United States of America·C Lo BiancoP Aebischer
Apr 30, 2003·The Journal of Biological Chemistry·Julie L WebbDavid C Rubinsztein
Aug 19, 2003·Human Genetics·Ornit Chiba-FalekRobert L Nussbaum
Nov 1, 2003·Science·A B SingletonK Gwinn-Hardy
Sep 29, 2004·Lancet·Marie-Christine Chartier-HarlinAlain Destée
Dec 4, 2004·Proceedings of the National Academy of Sciences of the United States of America·Christophe Lo BiancoPatrick Aebischer
Feb 8, 2006·Biochemical and Biophysical Research Communications·Hiromi Hayashita-KinohHideki Mochizuki
Jun 24, 2006·Movement Disorders : Official Journal of the Movement Disorder Society·Ornit Chiba-FalekRobert L Nussbaum
Jul 11, 2006·Journal of Cellular Physiology·Ayumi SatoMami Noda
Jul 14, 2006·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Yan Leng, De-Maw Chuang
Mar 29, 2008·Science·Job Dekker
Jun 23, 2009·Current Protein & Peptide Science·Alison L McCormack, Donato A Di Monte
Mar 5, 2010·The Journal of Biological Chemistry·Sally K MakDonato A Di Monte
May 7, 2010·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Ahmad JowaedUllrich Wüllner
May 12, 2010·Proceedings of the National Academy of Sciences of the United States of America·Youren TongJie Shen
Nov 6, 2010·Molecular Neurodegeneration·Preeti J KhandelwalCharbel Eh Moussa
Feb 10, 2011·Neurobiology of Disease·Lydia Alvarez-ErvitiJ Mark Cooper
Sep 3, 2011·PloS One·Byung Rho Lee, Tetsu Kamitani
Sep 9, 2011·The Journal of Biological Chemistry·Sharan PaulSita Reddy

❮ Previous
Next ❯

Citations

May 17, 2018·Cell Transplantation·Hueng-Chuen FanHorng-Jyh Harn
Oct 6, 2016·Archives of Toxicology·Alejandro RomeroMaría-Aránzazu Martínez
Mar 18, 2019·Journal of Neuroimmune Pharmacology : the Official Journal of the Society on NeuroImmune Pharmacology·Jie LuoAnumantha G Kanthasamy

❮ Previous
Next ❯

Datasets Mentioned

BETA
ENSG00000145335

Methods Mentioned

BETA
PCR
Assay
transfection

Related Concepts

Related Feeds

Basal Ganglia

Basal Ganglia are a group of subcortical nuclei in the brain associated with control of voluntary motor movements, procedural and habit learning, emotion, and cognition. Here is the latest research.

Antisense Oligonucleotides: ND

This feed focuses on antisense oligonucleotide therapies such as Inotersen, Nusinursen, and Patisiran, in neurodegenerative diseases including amyotrophic lateral sclerosis.

Alpha-Synuclein Aggregation

Alpha-synucleins are small proteins that are believed to restrict the mobility of synpatic vesicles and inhibit neurotransmitter release. Aggregation of these proteins have been linked to several types of neurodegenerative diseases including dementia with Lewy bodies and Parkinson's disease. Here is the latest research on α-synuclein aggregation.

CREs: Gene & Cell Therapy

Gene and cell therapy advances have shown promising outcomes for several diseases. The role of cis-regulatory elements (CREs) is crucial in the design of gene therapy vectors. Here is the latest research on CREs in gene and cell therapy.

Alzheimer's Disease: RNA Regulation

RNA regulation involves several mechanisms that are used by cells to decrease or increase the production of RNA. Disruption of RNA regulatory processes has been associated with Alzheimer's disease (AD). Here are the latest discoveries pertaining to RNA regulation and AD.

Attention Disorders

Attention is involved in all cognitive activities, and attention disorders are reported in patients with various neurological diseases. Here are the latest discoveries pertaining to attention disorders.

Alzheimer's Disease: Transcription

Impaired transcription is associated with the pathogenesis and progression of conditions such as Alzheimer's disease (AD). Here are the latest discoveries pertaining to transcription and AD.

Alzheimer's Disease: Genetics

Alzheimer's disease is a neurodegenerative disease. Discover genetic and epigenetic aspects of Alzheimer’s disease, including genetic markers and genomic structural variations with this feed.

Alpha-Synuclein Aggregation (MDS)

Alpha-synucleins are small proteins that are believed to restrict the mobility of synpatic vesicles and inhibit neurotransmitter release. Aggregation of these proteins have been linked to several types of neurodegenerative diseases including dementia with Lewy bodies and Parkinson's disease. Here is the latest research on α-synuclein aggregation.