초록 ▼

1단계 4년간의 노인성 뇌질환 형질전환 동물모델 개발 사업을 통해 렌티바이러스 벡터 시스템 개발, CRISPR/CAS 9 이용 유전자 적중 벡터 구축 및 유효성 확인 체계 구축, 퇴행성 뇌질환 유전자 과발현 세포주 구축, 고효율 난배란 유도 및 정액 채취 최적화 시스템 구축, 난세포질내 정자주입법 (ICSI)을 통한 원숭이 수정란 생산 기술 확립, 렌티바이러스 및 유전자 가위 활용 형질전환 수정란 개발 기술 확립 등을 통해 국내 최초로 뇌질환 형질전환 영장류 모델 및 유전자 적중 원숭이 생산 체계의 구축을 완료하였음. 이러한 기술을

1단계 4년간의 노인성 뇌질환 형질전환 동물모델 개발 사업을 통해 렌티바이러스 벡터 시스템 개발, CRISPR/CAS 9 이용 유전자 적중 벡터 구축 및 유효성 확인 체계 구축, 퇴행성 뇌질환 유전자 과발현 세포주 구축, 고효율 난배란 유도 및 정액 채취 최적화 시스템 구축, 난세포질내 정자주입법 (ICSI)을 통한 원숭이 수정란 생산 기술 확립, 렌티바이러스 및 유전자 가위 활용 형질전환 수정란 개발 기술 확립 등을 통해 국내 최초로 뇌질환 형질전환 영장류 모델 및 유전자 적중 원숭이 생산 체계의 구축을 완료하였음. 이러한 기술을 바탕으로 Tyrosinase유전자 KO 알비노 원숭이 사산개체생산에 성공하였으며, GFP 발현 형질전환 원숭이 생산에 성공하였음. 약물투여 비수술적 방법을 통한 유발퇴행성 중추신경계 질환 영장류 모델 개발을 수행하여 알츠하이머형 치매모델 제작 기술을 확립하였음. 알츠하이머형 치매모델의 제작은 Intracerebroventricular administration of streptozotocin 방법 및 Aβ oligomer의 도입에 의해서 제작하였음. 영장류 질환모델의 병리학적, 행동학적 및 영상진단학적 질환평가기술을 개발하여, 손기능 자동검사 기술, 인지기능 검사체계개발, 뇌조직 절단기 개발 등 다양한 기술 개발에 성공하여 7가지 관련 기술을 특허 출원하였으며, 1가지 기술을 특허 등록하였음. 영장류센터에서 자연적으로 노화가 진행되고 있는 아프리카녹색원숭이의 혈액을 이용하여 Genome, Transcriptome, 에피지놈 데이터를 수집하여 분석을 진행하였으며, 2단계 연구에 활용될 노화 유전체 데이터를 확보하였음. 1단계 4년동안 영장류 질화모델 제작, 영장류 Tg, KO생산 기술 개발 등 국내 최초의 다양한 기술 및 인프라를 확립하였으며, 국제학술지에 44편의 연구 논문을 발표하였음.

(출처 : 요약서 3p)

Abstract ▼

Ⅳ. Research Development Results (2014)
1. Development of transgenic technology and production of transformants for the development of degenerative brain disease primate models
- Establishment of monkey embryos microinjection conditions using the GFP-expressing lentivirus
- Construction and

Ⅳ. Research Development Results (2014)
1. Development of transgenic technology and production of transformants for the development of degenerative brain disease primate models
- Establishment of monkey embryos microinjection conditions using the GFP-expressing lentivirus
- Construction and validation of mutant DJ-1 overexpression lentiviral vector
- Monkey tyrosinase gene isolation and knockout RGEN design

2. Development of degenerative central nervous system disease primate model through drug administration and non-surgical methods
- Development of Alzheimer's Disease (AD) induction technology in primates using ICV-STZ
- Development of degenerative central nervous system impaired primate disease model

3. Development of disease assessment techniques including pathological, behavioral and diagnostic imaging for primate disease models
- Development of pathological and behavioral evaluation technology in AD disease model
- Construction of acquisition system for brain imaging of primate brain disease model
- Systematization of qualitative and quantitative evaluation in acquired brain image

4. Development of aging biomarker and useful gene discovery technology using the aging primate
- Secure an aging primate genome sequence through NGS analysis
- Secure an aging primate transcriptome sequence through NGS analysis

5. Development of degenerative brain disease/arthritis non-primate animal models
- Identification of chondrocyte property changes and construction of regulation method in inflammatory conditions
- Establishment of homeostatic regulation method of injured cartilage
- Development of degenerative neurological disease animal production technology
- Cognitive disorders - Dementia disease animal analysis
- Analysis of gene expression through microarray
- Analysis of gene biomarker
- construction of stabilization of replication efficiency
- Identification of molecular functions and mechanisms of DJ-1 in existing Parkinson's disease model animals
- Secure a phosphatase gene related to degenerative disease
- Establishment of phosphatase expression conditions
- Mass expression and purification of phosphatase

Ⅳ. Research Development Results (2015)
1. Development of transgenic technology and production of transformants for the development of degenerative brain disease primate models
- Establishment of transgenic embryo production technology through microinjection of mutant DJ-1 lentivirus
- Preparation of surrogate mother and establishment of embryo transfer technology
- Establishment of DJ-1 overexpressed high efficient vector and lentivirus system
- Neuronal cell mitochondrial morphological changes and finding of apoptosis regulation factor

2. Development of degenerative central nervous system disease primate model through drug administration and non-surgical methods
- Establishment of Alzheimer's Disease (AD) induction technology in primates using ICV-STZ
- Development of degenerative central nervous system impaired primate disease model

3. Development of disease assessment techniques including pathological, behavioral and diagnostic imaging for primate disease models
- Development of pathological and behavioral evaluation technology in AD disease model
- Construction of acquisition system for brain imaging of primate brain disease model
- Systematization of qualitative and quantitative evaluation in acquired brain image

4. Development of aging biomarker and useful gene discovery technology using the aging primate
- Securment of aging primates gene information by integrating genome sequence and transcriptome sequence
- Mapping aging primates methylome

5. Development of degenerative brain disease/arthritis non-primate animal models
- Construction of rodent animal model (MCAo)
- Finding of neural circuit damage mechanism in animal model
- Development of degenerative neurological disease animal production technology
- Cloning of biomarker candidates
- Development of biomarker candidates-inserted stem cell
- In vitro efficacy evaluation of candidate gene-inserted stem cells
- Production of cloned transgenic dog
- Securement of large quantity of phosphatase
- Construction of enzyme activity method
- Screening of natural products-derived compound libraries

Ⅳ. Research Development Results (2016)
1. Development of transgenic technology and production of transformants for the development of degenerative brain disease primate models
- Establishment of transformant production technology through microinjection of mutant DJ-1 lentivirus
- Establishment of tyrosinase knockout monkey embryo production technology through RGEN microinjection
- Production of CRISPR / Cas9-VPS35 vector
- Production of Aβ42 using recombinant oligomers
- Construction of mutant-type expression vector system for the Alzheimer's disease-inducing gene (APP / PS1)
- Establishment of overexpression vector system for Parkinson's disease gene (DJ-1, PINK1, Parkin)
- Establishment of oxidative stress and autophagy-specific response gene expression vector system and neuronal cell line in each organelle
- Development of production technology for transgenic primates
- Development of technology for optimal transgenic primate embryo production
- Development of a medium size animal model to identify disease potential of transgenic primates
- EMT and cell migration mechanism analysis of primate-derived cells and human-derived cancer cells
- Deduction of autophagy control mechanism in primate-derived cells

2. Development of degenerative central nervous system disease primate model through drug administration and non-surgical methods
- Production of primate Alzheimer's disease (AD) model using ICV-STZ
- Selective muscle degeneration experiment by mixture of Botox and Matrigel
- Verification of spinal disc degeneration following weakening of the spinal and back muscles
- Development of program for analysis of 3-D experimental animal behavior change

3. Development of disease assessment techniques including pathological, behavioral and diagnostic imaging for primate disease models
- Development of pathological and behavioral evaluation technology in AD disease model
- Construction of acquisition system for brain imaging of primate brain disease model
- Systematization of qualitative and quantitative evaluation in acquired brain image

4. Development of aging biomarker and useful gene discovery technology using the aging primate
- Development of biomarker through construction of database for analysis of primate genetic information, and finding useful genes
- Epigenome information analysis and marker development of aging primates

5. Development of degenerative brain disease/arthritis non-primate animal models
- Identification of the role of Pin1 / DAPK1 in ischemic brain injury
- Establishment of brain tissue culture model based on rodent model (Ex vivo)
- Identification of phosphorylation and protein stability changes of TrCP by controlling Pin1 / DAPK1 in established primate cell-lines
- Development of diabetes-mediated dementia model
- Analysis of blood glucose and cognitive impairment behavior
- Analysis of animal models characterization
- Synthesis of beta amyloid fluorescent probe (two or more)
- Optimization of beta amyloid precipitation detection system
- Candidate substance search for inhibition of beta amyloid deposition

Ⅳ. Research Development Results (2017)
1. Development of transgenic technology and production of transformants for the development of degenerative brain disease primate models
- Establishment of transformant production technology through microinjection of mutant DJ-1 lentivirus
- Establishment of tyrosinase knockout monkey production technology through RGEN microinjection
- Production of CRISPR / Cas9-VPS35 vector
- Construction of degenerative brain disease-induced gene transformation vector system and expression verification in primate-derived neuronal cells
- Identification of mechanism of chronic degenerative disease in transgenic primate brain disease model
- Construction of molecular mechanism analysis system for degenerative brain disease-induced genes in primate-derived neuronal cells

2. Development of degenerative central nervous system disease primate model through drug administration and non-surgical methods
- Production of primate Alzheimer's disease(AD) model using ICV-STZ & Aβ oligomer
- Development of degenerative central nervous system impaired primate disease model

3. Development of disease assessment techniques including pathological, behavioral and diagnostic imaging for primate disease models
- Development of pathological and behavioral evaluation technology in AD disease model
- Construction of acquisition system for brain imaging of primate brain disease model
- Systematization of qualitative and quantitative evaluation in acquired brain image

4. Development of aging biomarker and useful gene discovery technology using the aging primate
- Development of biomarker through construction of database for analysis of primate genetic information, and finding useful genes
- Epigenome information analysis and marker development of aging primates

5. Development of degenerative brain disease/arthritis non-primate animal models
- Construction of research base on analysis of brain cognitive function regulation and signal transmission system through establishment of ex vivo system using primate brain cell and tissue
- Pathophysiological analysis using rodent-induced model for development of optimal primate vascular dementia disease model
- Study on the mechanism of amyloid beta peptide plaque formation and development of inhibitor detection system using primate-derived cells

(출처 : SUMMARY 12p)

목차 Contents

• 표지 ... 1
• 제 출 문 ... 2
• 보고서 요약서 ... 3
• 요 약 문 ... 4
• SUMMARY ... 12
• CONTENTS ... 21
• 목차 ... 22
• 제 1 장 연구개발과제의 개요 ... 23
• 제 1 절 연구개발의 목적 ... 23
• 제 2 절 연구개발의 필요성 ... 23
• 제 2 장 국내외 기술개발 현황 ... 34
• 제 1 절 국내·외 연구 개발 현황 및 연구결과의 국내·외 기술개발 현황에서 차지하는 위치 ... 34
• 제 3 장 연구개발수행 내용 및 결과 ... 47
• 제 1 절 2014년도 연구개발 수행 내용 및 결과 ... 47
• 제 2 절 2015년도 연구개발 수행 내용 및 결과 ... 113
• 제 3 절 2016년도 연구개발 수행 내용 및 결과 ... 136
• 제 4 절 2017년도 연구개발 수행 내용 및 결과 ... 204
• 제 4 장 목표달성도 및 관련분야에의 기여도 ... 332
• 제 1 절 목표 달성도 ... 332
• 제 2 절 관련분야 기여도 ... 344
• 제 5 장 연구개발결과의 활용계획 ... 346
• 제 6 장 연구개발과정에서 수집한 해외과학기술정보 ... 348
• 제 1 절 형질전환/복제 기술 이용 영장류 질환모델 연구 동향 ... 348
• 제 7 장 연구시설 및 장비 현황 ... 355
• 제 8 장 참고문헌 ... 356
• 끝페이지 ... 359