{"id":{"repo_id":"uiuc","oai_identifier":"oai:www.ideals.illinois.edu:2142/106499"},"canonical_url":"https://search.dev.ndltd.org/etd/uiuc/oai:www.ideals.illinois.edu:2142/106499","repository":{"repo_id":"uiuc","name":"University of Illinois - Urbana-Champaign","base_url":"https://www.ideals.illinois.edu/oai-pmh"},"display":{"title":"Genetic correction of Parkinson’s disease using CRISPR-SKIP","abstract":"Parkinson’s Disease (PD) is the second most common neurodegenerative disease after Alzheimer’s disease. It affects 1-2% of the total world population with more than 500,000 patients in the US. PD is a progressive disorder that selectively affects the dopaminergic neurons in the substantia nigra of brain. Since dopamine is the neurotransmitter responsible for controlling balance and movement of the body, patients affected by PD experience symptoms such as tremors, bradykinesia, limb rigidity, gait and balance problems. A common pathogenic mechanism in both familial and sporadic forms of PD is the accumulation of the protein α-synuclein in the brain basal ganglia. SNCA, the gene encoding α-synuclein, when mutated or overexpressed causes misfolding of α-synuclein, which forms toxic aggregates known as Lewy bodies, a characteristic pathological finding in PD. There is no cure for PD and the only treatment available is symptomatic therapy. Therefore, there is a critical need to engineer novel approaches for treating PD. A potential strategy to prevent neurotoxicity in PD is to force the affected cells to produce less toxic isoforms of SNCA by skipping the exons that encode for the domains responsible for aggregation. Thus, one potential approach to treat PD is skipping exon 3 in SNCA, which has been shown to decrease α-synuclein aggregation. We have recently demonstrated that CRISPR-Cas9 base editors can be used to induce exon skipping by mutating the conserved “AG” dinucleotide within the splice acceptor site preceding each exon. Here we demonstrate skipping of exon 3 in the SNCA gene in mammalian cells in culture by A>G and C>T base editors, which decreased SNCA aggregation. We also demonstrate that skipping of SNCA exon 3 can be accomplished using split base editors that are compatible with in vivo delivery. Overall, these results demonstrate development of a novel gene therapy to treat PD.","abstract_html":"Parkinson’s Disease (PD) is the second most common neurodegenerative disease after Alzheimer’s disease. It affects 1-2% of the total world population with more than 500,000 patients in the US. PD is a progressive disorder that selectively affects the dopaminergic neurons in the substantia nigra of brain. Since dopamine is the neurotransmitter responsible for controlling balance and movement of the body, patients affected by PD experience symptoms such as tremors, bradykinesia, limb rigidity, gait and balance problems. A common pathogenic mechanism in both familial and sporadic forms of PD is the accumulation of the protein α-synuclein in the brain basal ganglia. SNCA, the gene encoding α-synuclein, when mutated or overexpressed causes misfolding of α-synuclein, which forms toxic aggregates known as Lewy bodies, a characteristic pathological finding in PD. There is no cure for PD and the only treatment available is symptomatic therapy. Therefore, there is a critical need to engineer novel approaches for treating PD. A potential strategy to prevent neurotoxicity in PD is to force the affected cells to produce less toxic isoforms of SNCA by skipping the exons that encode for the domains responsible for aggregation. Thus, one potential approach to treat PD is skipping exon 3 in SNCA, which has been shown to decrease α-synuclein aggregation. We have recently demonstrated that CRISPR-Cas9 base editors can be used to induce exon skipping by mutating the conserved “AG” dinucleotide within the splice acceptor site preceding each exon. Here we demonstrate skipping of exon 3 in the SNCA gene in mammalian cells in culture by A>G and C>T base editors, which decreased SNCA aggregation. We also demonstrate that skipping of SNCA exon 3 can be accomplished using split base editors that are compatible with in vivo delivery. Overall, these results demonstrate development of a novel gene therapy to treat PD.","abstract_has_math":false,"creators":["Manandhar, Sony"],"institution":"University of Illinois at Urbana-Champaign","degree_name":"M.S.","degree_level":"Thesis","degree_discipline":"Bioengineering","degree_department":null,"school":null,"contributors":["Perez-Pinera, Pablo"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2020,"date_issued":"2020-03-02T22:38:59Z","date_published":"2020-03-02T22:38:59Z","updated_at":"2026-07-22T22:24:47Z","subjects":["CRISPR-SKIP, Parkinson's Disease, SNCA, α-synuclein, Aggregation"],"languages":["en"],"rights":["Copyright 2019 Sony Manandhar"],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"http://hdl.handle.net/2142/106499","outbound_label":"Handle","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Perez-Pinera, Pablo"]},{"key":"dc:creator","label":"Author","values":["Manandhar, Sony"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2020-03-02T22:38:59Z","2022-03-03T10:15:08Z","2019-12-11","2019-12"]},{"key":"dc:type","label":"Dc Type","values":["text"]},{"key":"thesis:degree_discipline","label":"Discipline","values":["Bioengineering"]},{"key":"thesis:degree_level","label":"Degree Level","values":["Thesis"]},{"key":"thesis:degree_name","label":"Degree Name","values":["M.S."]},{"key":"thesis:institution_name","label":"Thesis Institution Name","values":["University of Illinois at Urbana-Champaign"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["CRISPR-SKIP, Parkinson's Disease, SNCA, α-synuclein, Aggregation"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["en"]},{"key":"dc:rights","label":"Dc Rights","values":["Copyright 2019 Sony Manandhar"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["http://hdl.handle.net/2142/106499"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["Parkinson’s Disease (PD) is the second most common neurodegenerative disease after Alzheimer’s disease. It affects 1-2% of the total world population with more than 500,000 patients in the US. PD is a progressive disorder that selectively affects the dopaminergic neurons in the substantia nigra of brain. Since dopamine is the neurotransmitter responsible for controlling balance and movement of the body, patients affected by PD experience symptoms such as tremors, bradykinesia, limb rigidity, gait and balance problems. A common pathogenic mechanism in both familial and sporadic forms of PD is the accumulation of the protein α-synuclein in the brain basal ganglia. SNCA, the gene encoding α-synuclein, when mutated or overexpressed causes misfolding of α-synuclein, which forms toxic aggregates known as Lewy bodies, a characteristic pathological finding in PD. There is no cure for PD and the only treatment available is symptomatic therapy. Therefore, there is a critical need to engineer novel approaches for treating PD. A potential strategy to prevent neurotoxicity in PD is to force the affected cells to produce less toxic isoforms of SNCA by skipping the exons that encode for the domains responsible for aggregation. Thus, one potential approach to treat PD is skipping exon 3 in SNCA, which has been shown to decrease α-synuclein aggregation. We have recently demonstrated that CRISPR-Cas9 base editors can be used to induce exon skipping by mutating the conserved “AG” dinucleotide within the splice acceptor site preceding each exon. Here we demonstrate skipping of exon 3 in the SNCA gene in mammalian cells in culture by A>G and C>T base editors, which decreased SNCA aggregation. We also demonstrate that skipping of SNCA exon 3 can be accomplished using split base editors that are compatible with in vivo delivery. Overall, these results demonstrate development of a novel gene therapy to treat PD.","Submission published under a 24 month embargo labeled 'Closed Access', the embargo will last until 2021-12-01","The student, Sony Manandhar, accepted the attached license on 2019-12-11 at 13:14.","The student, Sony Manandhar, submitted this Thesis for approval on 2019-12-11 at 13:15.","This Thesis was approved for publication on 2019-12-11 at 16:06.","DSpace SAF Submission Ingestion Package generated from Vireo submission #14791 on 2020-02-28 at 17:38:23","Made available in DSpace on 2020-03-02T22:38:59Z (GMT). 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It affects 1-2% of the total world population with more than 500,000 patients in the US. PD is a progressive disorder that selectively affects the dopaminergic neurons in the substantia nigra of brain. Since dopamine is the neurotransmitter responsible for controlling balance and movement of the body, patients affected by PD experience symptoms such as tremors, bradykinesia, limb rigidity, gait and balance problems. A common pathogenic mechanism in both familial and sporadic forms of PD is the accumulation of the protein α-synuclein in the brain basal ganglia. SNCA, the gene encoding α-synuclein, when mutated or overexpressed causes misfolding of α-synuclein, which forms toxic aggregates known as Lewy bodies, a characteristic pathological finding in PD. There is no cure for PD and the only treatment available is symptomatic therapy. Therefore, there is a critical need to engineer novel approaches for treating PD. A potential strategy to prevent neurotoxicity in PD is to force the affected cells to produce less toxic isoforms of SNCA by skipping the exons that encode for the domains responsible for aggregation. Thus, one potential approach to treat PD is skipping exon 3 in SNCA, which has been shown to decrease α-synuclein aggregation. We have recently demonstrated that CRISPR-Cas9 base editors can be used to induce exon skipping by mutating the conserved “AG” dinucleotide within the splice acceptor site preceding each exon. Here we demonstrate skipping of exon 3 in the SNCA gene in mammalian cells in culture by A>G and C>T base editors, which decreased SNCA aggregation. We also demonstrate that skipping of SNCA exon 3 can be accomplished using split base editors that are compatible with in vivo delivery. Overall, these results demonstrate development of a novel gene therapy to treat PD.","Submission published under a 24 month embargo labeled 'Closed Access', the embargo will last until 2021-12-01","The student, Sony Manandhar, accepted the attached license on 2019-12-11 at 13:14.","The student, Sony Manandhar, submitted this Thesis for approval on 2019-12-11 at 13:15.","This Thesis was approved for publication on 2019-12-11 at 16:06.","DSpace SAF Submission Ingestion Package generated from Vireo submission #14791 on 2020-02-28 at 17:38:23","Made available in DSpace on 2020-03-02T22:38:59Z (GMT). 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