초록 ▼

○ 핵심은 유전자 및 게놈을 저비용, 초고속으로 생산하는 것: 생명과학기술의 패러다임이 기존의 개별 유전자만을 연구하는 것으로부터, 생명체의 게놈 전체를 분석 및 조작하는 것으로 변화하고 있음. 현재 이 흐름을 막는 가장 큰 문제는 게놈 수준의 DNA을 생산하는 데 천문학 적인 비용과 노력이 든다는데 있음. 본 연구단에서는 이 문제를, 4가지 핵심 기술요소를 개발하고 각 요소들을 융합하여 독자적인 원천기반기술을 확보함으로써 극복함.
○ 초고밀도, 초고순도의 올리고뉴클레오타이드 어레이 칩 제조 기술: 게놈 수준의 DNA의 생산

○ 핵심은 유전자 및 게놈을 저비용, 초고속으로 생산하는 것: 생명과학기술의 패러다임이 기존의 개별 유전자만을 연구하는 것으로부터, 생명체의 게놈 전체를 분석 및 조작하는 것으로 변화하고 있음. 현재 이 흐름을 막는 가장 큰 문제는 게놈 수준의 DNA을 생산하는 데 천문학 적인 비용과 노력이 든다는데 있음. 본 연구단에서는 이 문제를, 4가지 핵심 기술요소를 개발하고 각 요소들을 융합하여 독자적인 원천기반기술을 확보함으로써 극복함.
○ 초고밀도, 초고순도의 올리고뉴클레오타이드 어레이 칩 제조 기술: 게놈 수준의 DNA의 생산 비용을 획기적으로 줄이기 위해서 어레이 칩 기반의 올리고뉴클레오타이드 합성은 필수적임. 나노광학적인 방법을 적용, 단분자 기반의 초고밀도, 초고순도의 올리고뉴클레오타이드 어레이 칩 제조 기술을 개발함.
○ 나노-반도체 융합을 통한 차세대 염기서열 분석 기술: 게놈 수준의 DNA를 합성하기 위해서는 조립과정 전에 오류가 없는 올리고뉴클레오타이드를 선별하는 과정이 필수적임. 본 연구단에서는 나노-반도체 기반의 차세대 염기서열 분석기술을 독자적으로 개발하여 게놈 조립과정에서 발생할 수 있는 오류를 원천봉쇄함.
○ 나노/미세유체 기술을 이용한 DNA 농축 및 조립 기술: 게놈 수준의 DNA 생산 수율을 증가시키고 생산 비용을 줄이기 위해, 나노/미세유체 기술을 개발하여 미세유체관의 국소부분에 DNA를 농축시

(출처 : 보고서 요약서 3p)

Abstract ▼

Ⅳ. Results
Our research group has developed four foundation technologies for synthesizing genes in cheap and high throughput manner, which reduced the cost 100 times for synthesizing 1 MB-long gene.
We have secured the source technology (13 patent registrations) and also worked on the commerci

Ⅳ. Results
Our research group has developed four foundation technologies for synthesizing genes in cheap and high throughput manner, which reduced the cost 100 times for synthesizing 1 MB-long gene.
We have secured the source technology (13 patent registrations) and also worked on the commercialization( 7 technology transfers, products were released ).

High throughput retrieval of molecular clones using pulse laser-based nano optics
• Convergence of molecular cloning method using nano-optics and next-generation sequencing method allows the acquisition of 200bp targeted DNA 200 times cheaper than existing methods and also increases the productivity more than 100 times. We also successfully acquired the foundation technology, which resulted in patent registration (3), technology transfer (4 case) and publication in Nature communications.

Next generation sequencing and bead retrieval using C chip
We have developed a method of designing and manufacturing bio-sensor in 4K level using CMOS semiconductor process. We also developed electrochemical AC biasing method by interpreting the chemical reaction mechanisms of the antibodies in the antigen of blood or serum and on the surface of C-chip sensor in order to improve on the existing electric method. In addition, We have checked the idea to improving the sensor performance (sensitivity and affinity) by adapting the electrical methods (ex, pulse method). By introducing the two-electrode system (reference electrode-free) instead of the conventional three electrode system, the electrical potential of the electrolyte (including the bio-solution) can be stabilized. The enhancement of the sensitivity and the selectivity in biosensor is important factor to increase the accuracy of the sensor and achievement of the real application. The trials have been carried out to enhancing the sensitivity and the selectivity. the result of the research trials are published to the academic journals. (In the case of the C-chip, the ACEF effect by using AC voltage, in the case of the T-chip, the extraction of the tunneling current by using the pulse method.) We have achieved paper publication (international: SCI (4), (Lab on a chip and Journal of Applied Physics) non-SCI (1) ), invited talk - international (2) , conference - domestic (5) international (5), patent application - domestic (2) international (2) , patent registration - domestic (2) international (2) and text book (1)

Next Generation DNA Assembler
• Horizontal DNA assembly technology
We divided microchip oligos, composed of tens of thousands of oligonucleotides, into 16 pools, and the oligos in each pool have a common flanking sequences at both ends which is distinguishable to other pool. With this oligo pools, we developed pool to pool DNA assembly method by amplifying each of 16 pools, cutting out flanking sequences, and then assembling them in hierarchical manner.

• Ultra low-cost production technology of synthetic genes using Illumina next-generation sequencing technology.
We first developed a method to analyze each target sequences through multiple target loci amplify and assembly for sanger sequencing. However, we have identified that there was a large difference in scalability between sanger sequencing and Illumina next-generation sequencing.
Since then, we have developed error-free microchip oligo retriever method and its device, sequencing analysis method of synthesized gene using Illumina sequencing technology.
We have also developed a techniques such as flexible enzyme recognition site DNA cloning method, repetitive gene insertion method to bacterial genomic DNA, and genome engineering method using dsDNA. Through development process, we acquired 11 patent applications (5 Domestic / 6 International), 5 patent registration (5 Domestic), and 4 publications in 1 in Nature communications, 1 in Nucleic Acids Research, and 2 in Scientific Reports.

• High-throughput DNA synthesis technology using Molecular Inversion Probes
We developed a technique to synthesize sequences of more than a thousands target regions using molecular inversion probes that bind to target DNA in inverted form. As a result, we published 1 paper in Nucleic Acids Research.

• Multi-layer oligonucleotide design and synthesis technology for high-efficiency gene synthesis
We developed a new gene synthesis designing method with multi-layered microchip oligos and showed improved efficiency compared with common gene synthesis method. We successfully synthesized tens of large sized genes (>4kb) simultaneously. As a result, we acquired 3 patent applications (2 Domestic / 1 international) and published 1 paper in Nucleic Acids Research.

• A platform for sample preparation of gene library simultaneously with fast and inexpensive manner
The existing sanger sequencing method can read one DNA at a length of about 1,000 bp, but it was not suitable for analyzing hundreds or thousands of gene libraries. In order to analyze this through the Illumina sequencing technology, we implemented a transposon technique to cut each gene into a size suitable for the Illumina sequencing and attach the barcode sequence at each ends.

• Develop an analysis platform suitable for analyzing gene library from sequencing data at once
According to the technology at 5. every DNA fragments have common flanking sequence and barcode sequence at each ends. For convenient analysis, we developed a method that remove the common flanking sequence, sorting with barcode sequences, and assembly the remaining sequence data with de novo assembly method for gaining full gene length sequence. With this method, we submitted one paper to Bioinformatics.

Development of nanoliter-scale DNA reactor
• We have completed specimen concentration technique that can work on high buffer concentration and wide pH range. We also developed a technology that can concentrate both anions and cations, selectively. This is the first development in the world and related paper has been published on Lab-on-a-chip and a patent has been applied. Based on this technology, we have successfully concentrated DNA and DNA ligase in a very small volume simultaneously. Through this, we showed the possibility of error-free DNA synthesis using very small amount of DNA fragments. Also, we have transferred the high voltage control system design technology for specimen concentration micro fluidic chip (1 case) to a business enterprise.

(출처 : SUMMARY 16p)

목차 Contents

• 표지 ... 1
• 제 출 문 ... 2
• 보고서 요약서 ... 3
• 요 약 문 ... 4
• SUMMARY ... 13
• CONTENTS ... 23
• 목차 ... 24
• 제 1 장 연구개발과제의 개요 ... 25
• 제1절. 연구개발과제의 목적, 필요성 및 범위 ... 25
• 제2절. 대상 융합기술의 정의 및 개념 ... 28
• 제 2 장 국내외 기술개발 현황 ... 35
• 제 3 장 연구개발수행 내용 및 결과 ... 43
• 제1절. 이론적, 실험적 접근방법 ... 43
• 제2절. 연구내용 및 결과 ... 66
• 제 4 장 목표달성도 및 관련분야에의 기여도 ... 184
• 제1절. 연구개발목표의 달성도 ... 184
• 제2절. 관련분야의 기술발전에의 기여도 ... 197
• 제 5 장 연구개발결과의 활용계획 ... 200
• 제 6 장 연구개발과정에서 수집한 해외과학기술정보 ... 206
• 제 7 장 참고문헌 ... 210
• 끝페이지 ... 211