title: Lost region in amyloid precursor protein (APP) through TALEN-mediated genome editing alters mitochondrial morphology.
abstract: Alzheimer's disease (AD) is characterized by amyloid-β (Aβ) deposition in the brain. Aβ plaques are produced through sequential β/γ cleavage of amyloid precursor protein (APP), of which there are three main APP isoforms: APP695, APP751 and APP770. KPI-APPs (APP751 and APP770) are known to be elevated in AD, but the reason remains unclear. Transcription activator-like (TAL) effector nucleases (TALENs) induce mutations with high efficiency at specific genomic loci, and it is thus possible to knock out specific regions using TALENs. In this study, we designed and expressed TALENs specific for the C-terminus of APP in HeLa cells, in which KPI-APPs are predominantly expressed. The KPI-APP mutants lack a 12-aa region that encompasses a 5-aa trans-membrane (TM) region and 7-aa juxta-membrane (JM) region. The mutated KPI-APPs exhibited decreased mitochondrial localization. In addition, mitochondrial morphology was altered, resulting in an increase in spherical mitochondria in the mutant cells through the disruption of the balance between fission and fusion. Mitochondrial dysfunction, including decreased ATP levels, disrupted mitochondrial membrane potential, increased ROS generation and impaired mitochondrial dehydrogenase activity, was also found. These results suggest that specific regions of KPI-APPs are important for mitochondrial localization and function.
authors.affiliation: Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education, School of Pharmaceutical Sciences, and Medical Research Institute, Wuhan University, Wuhan 430071, China.
authors.forename: Yajie
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education, School of Pharmaceutical Sciences, and Medical Research Institute, Wuhan University, Wuhan 430071, China.|Taihe Hospital, Hubei University of Medicine, Shiyan, Hubei, China.
authors.forename: Fengyi
authors.initials: F
authors.lastname: Wu
authors.affiliation: Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education, School of Pharmaceutical Sciences, and Medical Research Institute, Wuhan University, Wuhan 430071, China.
authors.forename: Haining
authors.initials: H
authors.lastname: Pan
authors.affiliation: Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education, School of Pharmaceutical Sciences, and Medical Research Institute, Wuhan University, Wuhan 430071, China.
authors.forename: Wenzhong
authors.initials: W
authors.lastname: Zheng
authors.affiliation: Taihe Hospital, Hubei University of Medicine, Shiyan, Hubei, China.
authors.forename: Chi
authors.initials: C
authors.lastname: Feng
authors.affiliation: Taihe Hospital, Hubei University of Medicine, Shiyan, Hubei, China.
authors.forename: Yunfu
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education, School of Pharmaceutical Sciences, and Medical Research Institute, Wuhan University, Wuhan 430071, China.
authors.forename: Zixin
authors.initials: Z
authors.lastname: Deng
authors.affiliation: Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education, School of Pharmaceutical Sciences, and Medical Research Institute, Wuhan University, Wuhan 430071, China.
authors.forename: Lianrong
authors.initials: L
authors.lastname: Wang
authors.affiliation: Taihe Hospital, Hubei University of Medicine, Shiyan, Hubei, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Luo
authors.affiliation: Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education, School of Pharmaceutical Sciences, and Medical Research Institute, Wuhan University, Wuhan 430071, China.
authors.forename: Shi
authors.initials: S
authors.lastname: Chen
mesh_terms: D000595:Amino Acid Sequence; D016564:Amyloid beta-Protein Precursor / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D001665:Binding Sites; D002460:Cell Line; D017124:Conserved Sequence; D000072669:Gene Editing; D006801:Humans; D008928:Mitochondria / Q000235:genetics* / Q000378:metabolism*; D063154:Mitochondrial Dynamics / Q000235:genetics; D009154:Mutation; D059372:Nucleotide Motifs; D011485:Protein Binding; D000069896:Transcription Activator-Like Effector Nucleases
chemical_list: D016564:Amyloid beta-Protein Precursor; D000069896:Transcription Activator-Like Effector Nucleases