CRISPR-Cas9 Gene Editing Protects from the A53T-SNCA Overexpression-Induced Pathology of Parkinson's Disease in Vivo

Hyung Ho Yoon; Sunghyeok Ye; Sunhwa Lim; Ara Jo; Hawon Lee; Felix Hong; Seung Eun Lee; Soo Jin Oh; Na Rae Kim; Kyoungmi Kim; Bum-Joon Kim; Hyunjin Kim; Changjoon Lee; Min Ho Nam; Junseok W. Hur; Sang Ryong Jeon

doi:10.1089/crispr.2021.0025

CRISPR-Cas9 Gene Editing Protects from the A53T-SNCA Overexpression-Induced Pathology of Parkinson's Disease in Vivo

Hyung Ho Yoon, Sunghyeok Ye, Sunhwa Lim, Ara Jo, Hawon Lee, Felix Hong, Seung Eun Lee, Soo Jin Oh, Na Rae Kim, Kyoungmi Kim, Bum-Joon Kim, Hyunjin Kim, Changjoon Lee, Min Ho Nam, Junseok W. Hur, Sang Ryong Jeon

Department of Biomedical Sciences

Research output: Contribution to journal › Article › peer-review

23 Citations (Scopus)

Abstract

Mutations in specific genes, including synuclein alpha (SNCA) that encodes the α-synuclein protein, are known to be risk factors for sporadic Parkinson's disease (PD), as well as critical factors for familial PD. In particular, A53T-mutated SNCA (A53T-SNCA) is a well-studied familial pathologic mutation in PD. However, techniques for deletion of the mutated SNCA gene in vivo have not been developed. Here, we used the CRISPR-Cas9 system to delete A53T-SNCA in vitro as well as in vivo. Adeno-associated virus carrying SaCas9-KKH with a single-guide RNA targeting A53T-SNCA significantly reduced A53T-SNCA expression levels in vitro. Furthermore, we tested its therapeutic potential in vivo in a viral A53T-SNCA-overexpressing rat model of PD. Gene deletion of A53T-SNCA significantly rescued the overexpression of α-synuclein, reactive microgliosis, dopaminergic neurodegeneration, and parkinsonian motor symptoms. Our findings propose CRISPR-Cas9 system as a potential prevention strategy for A53T-SNCA-specific PD.

Original language	English
Pages (from-to)	95-108
Number of pages	14
Journal	CRISPR Journal
Volume	5
Issue number	1
DOIs	https://doi.org/10.1089/crispr.2021.0025
Publication status	Published - 2022 Feb

Bibliographical note

Funding Information:
This study was supported by grants from the Basic Science Research Program through the National Research Foundation (NRF) funded by the Korean Ministry of Education, Science and Technology (KR) (NRF-2017 R1D1A1B03035760, NRF-2019R1C1C1010602), Korea University, Republic of Korea (K1722461, K1809751, K2014091), and KU Medicine-KIST, Republic of Korea (O1902761) to J.W.H.; KIST institutional program (Project No. 2E30963) and NRF-2020M3E5D9079744 funded by the Ministry of Science and ICT of Korea to M.H.N.; and Asan Institute for Life Sciences Grant (2020IL0039) funded by the Asan Medical Center, Seoul, Republic of Korea, to S.R.J.

Publisher Copyright:
© Copyright 2022, Mary Ann Liebert, Inc., publishers 2022.

ASJC Scopus subject areas

Biotechnology
Genetics

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10.1089/crispr.2021.0025

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@article{7d54dbcafe9c44c6b9945600bf554100,

title = "CRISPR-Cas9 Gene Editing Protects from the A53T-SNCA Overexpression-Induced Pathology of Parkinson's Disease in Vivo",

abstract = "Mutations in specific genes, including synuclein alpha (SNCA) that encodes the α-synuclein protein, are known to be risk factors for sporadic Parkinson's disease (PD), as well as critical factors for familial PD. In particular, A53T-mutated SNCA (A53T-SNCA) is a well-studied familial pathologic mutation in PD. However, techniques for deletion of the mutated SNCA gene in vivo have not been developed. Here, we used the CRISPR-Cas9 system to delete A53T-SNCA in vitro as well as in vivo. Adeno-associated virus carrying SaCas9-KKH with a single-guide RNA targeting A53T-SNCA significantly reduced A53T-SNCA expression levels in vitro. Furthermore, we tested its therapeutic potential in vivo in a viral A53T-SNCA-overexpressing rat model of PD. Gene deletion of A53T-SNCA significantly rescued the overexpression of α-synuclein, reactive microgliosis, dopaminergic neurodegeneration, and parkinsonian motor symptoms. Our findings propose CRISPR-Cas9 system as a potential prevention strategy for A53T-SNCA-specific PD.",

author = "Yoon, {Hyung Ho} and Sunghyeok Ye and Sunhwa Lim and Ara Jo and Hawon Lee and Felix Hong and Lee, {Seung Eun} and Oh, {Soo Jin} and Kim, {Na Rae} and Kyoungmi Kim and Bum-Joon Kim and Hyunjin Kim and Changjoon Lee and Nam, {Min Ho} and Hur, {Junseok W.} and Jeon, {Sang Ryong}",

note = "Funding Information: This study was supported by grants from the Basic Science Research Program through the National Research Foundation (NRF) funded by the Korean Ministry of Education, Science and Technology (KR) (NRF-2017 R1D1A1B03035760, NRF-2019R1C1C1010602), Korea University, Republic of Korea (K1722461, K1809751, K2014091), and KU Medicine-KIST, Republic of Korea (O1902761) to J.W.H.; KIST institutional program (Project No. 2E30963) and NRF-2020M3E5D9079744 funded by the Ministry of Science and ICT of Korea to M.H.N.; and Asan Institute for Life Sciences Grant (2020IL0039) funded by the Asan Medical Center, Seoul, Republic of Korea, to S.R.J. Publisher Copyright: {\textcopyright} Copyright 2022, Mary Ann Liebert, Inc., publishers 2022.",

year = "2022",

month = feb,

doi = "10.1089/crispr.2021.0025",

language = "English",

volume = "5",

pages = "95--108",

journal = "CRISPR Journal",

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AU - Yoon, Hyung Ho

AU - Ye, Sunghyeok

AU - Lim, Sunhwa

AU - Jo, Ara

AU - Lee, Hawon

AU - Hong, Felix

AU - Lee, Seung Eun

AU - Oh, Soo Jin

AU - Kim, Na Rae

AU - Kim, Kyoungmi

AU - Kim, Bum-Joon

AU - Kim, Hyunjin

AU - Lee, Changjoon

AU - Nam, Min Ho

AU - Hur, Junseok W.

AU - Jeon, Sang Ryong

N1 - Funding Information: This study was supported by grants from the Basic Science Research Program through the National Research Foundation (NRF) funded by the Korean Ministry of Education, Science and Technology (KR) (NRF-2017 R1D1A1B03035760, NRF-2019R1C1C1010602), Korea University, Republic of Korea (K1722461, K1809751, K2014091), and KU Medicine-KIST, Republic of Korea (O1902761) to J.W.H.; KIST institutional program (Project No. 2E30963) and NRF-2020M3E5D9079744 funded by the Ministry of Science and ICT of Korea to M.H.N.; and Asan Institute for Life Sciences Grant (2020IL0039) funded by the Asan Medical Center, Seoul, Republic of Korea, to S.R.J. Publisher Copyright: © Copyright 2022, Mary Ann Liebert, Inc., publishers 2022.

PY - 2022/2

Y1 - 2022/2

N2 - Mutations in specific genes, including synuclein alpha (SNCA) that encodes the α-synuclein protein, are known to be risk factors for sporadic Parkinson's disease (PD), as well as critical factors for familial PD. In particular, A53T-mutated SNCA (A53T-SNCA) is a well-studied familial pathologic mutation in PD. However, techniques for deletion of the mutated SNCA gene in vivo have not been developed. Here, we used the CRISPR-Cas9 system to delete A53T-SNCA in vitro as well as in vivo. Adeno-associated virus carrying SaCas9-KKH with a single-guide RNA targeting A53T-SNCA significantly reduced A53T-SNCA expression levels in vitro. Furthermore, we tested its therapeutic potential in vivo in a viral A53T-SNCA-overexpressing rat model of PD. Gene deletion of A53T-SNCA significantly rescued the overexpression of α-synuclein, reactive microgliosis, dopaminergic neurodegeneration, and parkinsonian motor symptoms. Our findings propose CRISPR-Cas9 system as a potential prevention strategy for A53T-SNCA-specific PD.

AB - Mutations in specific genes, including synuclein alpha (SNCA) that encodes the α-synuclein protein, are known to be risk factors for sporadic Parkinson's disease (PD), as well as critical factors for familial PD. In particular, A53T-mutated SNCA (A53T-SNCA) is a well-studied familial pathologic mutation in PD. However, techniques for deletion of the mutated SNCA gene in vivo have not been developed. Here, we used the CRISPR-Cas9 system to delete A53T-SNCA in vitro as well as in vivo. Adeno-associated virus carrying SaCas9-KKH with a single-guide RNA targeting A53T-SNCA significantly reduced A53T-SNCA expression levels in vitro. Furthermore, we tested its therapeutic potential in vivo in a viral A53T-SNCA-overexpressing rat model of PD. Gene deletion of A53T-SNCA significantly rescued the overexpression of α-synuclein, reactive microgliosis, dopaminergic neurodegeneration, and parkinsonian motor symptoms. Our findings propose CRISPR-Cas9 system as a potential prevention strategy for A53T-SNCA-specific PD.

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ER -

CRISPR-Cas9 Gene Editing Protects from the A53T-SNCA Overexpression-Induced Pathology of Parkinson's Disease in Vivo

Abstract

Bibliographical note

ASJC Scopus subject areas

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