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Aptamer–nanoparticle complexes as powerful diagnostic and therapeutic tools
조헌호,반창일 생화학분자생물학회 2016 Experimental and molecular medicine Vol.48 No.-
Correct diagnosis and successful therapy are extremely important to enjoy a healthy life when suffering from a disease. To achieve these aims, various cutting-edge technologies have been designed and fabricated to diagnose and treat specific diseases. Among these technologies, aptamer–nanomaterial hybrids have received considerable attention from scientists and doctors because they have numerous advantages over other methods, such as good biocompatibility, low immunogenicity and controllable selectivity. In particular, aptamers, oligonucleic acids or peptides that bind to a specific target molecule, are regarded as outstanding biomolecules. In this review, several screening techniques for aptamers, also called systematic evolution of ligands by exponential enrichment (SELEX) methods, are introduced, and diagnostic and therapeutic aptamer applications are also presented. Furthermore, we describe diverse aptamer–nanomaterial conjugate designs and their applications for diagnosis and therapy.
Highly sensitive and selective in vitro diagnostics based on DNA probes and aptamers
조헌호,이성환,반창일 한국구조생물학회 2015 Biodesign Vol.3 No.1
Medical diagnosis is very important and essential for maintaining a healthy life. In vitro diagnostics have recently been afocus of scientists and researchers because they have a number of merits over other diagnostic methods. In particular,nucleic acid-based diagnostic methods are powerful and promising techniques. In this review, various types of nucleicacid-based in vitro diagnostics are introduced. These methods can be categorized into three groups according to theiranalytical approaches. In addition, aptamer-based diagnostic methods are covered in greater detail because aptamers arepromising materials for diverse areas, not only as alternatives to antibodies but also as the core components of analyticalequipment. It is expected that in vitro diagnostics based on DNA probes and aptamers will become a valuable platformencompassing all types of diseases.
Chan Il Chang,강혜숙,반창일,김소연,Dong-ki Lee 한국분자세포생물학회 2009 Molecules and cells Vol.27 No.6
Chemically synthesized small interfering RNAs (siRNAs) can specifically knock-down expression of target genes via RNA interference (RNAi) pathway. To date, the length of synthetic siRNA duplex has been strictly maintained less than 30 bp, because an early study suggested that double-stranded RNAs (dsRNAs) longer than 30 bp could not trigger specific gene silencing due to the induction of non-specific antiviral interferon responses. Contrary to the current belief, here we show that synthetic dsRNA as long as 38 bp can result in specific target gene silencing without non-specific antiviral responses. Using this longer duplex structure, we have generated dsRNAs, which can simultaneously knock-down expression of two target genes (termed as dual-target siRNAs or dsiRNAs). Our results thus demonstrate the structural flexibility of gene silencing siRNAs, and provide a starting point to construct multifunctional RNA structures. The dsiRNAs could be utilized to develop a novel therapeutic gene silencing strategy against diseases with multiple gene alternations such as viral infection and cancer.
Crystal Structures of the Two Isomorphous A-DNA Decamers d(GTACGCGTAC) and d(GGCCGCGGCC)
김태균,권택훈,정혜선,구자강,Muttaiya Sundaralingam,반창일 대한화학회 2006 Bulletin of the Korean Chemical Society Vol.27 No.4
To study the effect of sequence on DNA structure, the two decamer crystal structures one alternating, d(GTACGCGTAC), and the other non-alternating, d(GGCCGCGGCC), were solved. Crystals of both decamers belong to the hexagonal space group P6122, with one strand in the asymmetric unit. The unit cell constants of the alternating decamer are a = b = 39.26 Å, c = 77.70 Å. The structure was refined with 1,828 reflections from 8.0 to 2.0 Å resolution to an R value of 21.3% with all DNA atoms and 63 water molecules. The isomorphous non-alternating decamer had unit cell dimensions of a = b = 39.05 Å, c = 82.15 Å. The structure was refined with 2,423 reflections from 8.0 to 2.0 Å resolution to a final R value of 22.2% for all DNA atoms and 65 water molecules. Although the average helical parameters of the decamers are typical of A-DNAs, there are some minor differences between them. The helical twist, rise, x-displacement, inclination and roll alternate in the alternating decamer, but do not in the non-alternating decamer. The backbone conformations in both structures show some differences; the residue G(7) of the alternating decamer is trans for a and g while the trans conformations are observed at the residue G(8) of the non-alternating decamer.
P-44Mismatched DNA 검출을 위한 단백질 고정화 기술의 개발
김기범,정우석,권낙현,박덕수,심윤보,반창일 한국화학공학회 2007 화학공학의이론과응용 Vol.10 No.2
본 연구는 mismatched DNA을 감지하기 위한 SH-SAW(Shear Horizontal Surface Acoustic Wave) 센서를 개발하기 위하여 mismatched DNA를 감지하기 위한 액상의 단백질을 Au이 증착되어 있는 QCM(Quartz Crystal Microbalance) 전극에 고정화하는 방법에 대하여 연구하였다. Au이 증착되어 있는 QCM전극 표면 위에 NTA 용액을 SAM(self assembled monolayer)을 형성시켰으며 MutS 단백질을 고정하기 위하여 NTA 단분자 표면에 촉매 역활을 하는 EDC를 고정한 후 mismatched DNA을 감지할 수 있는 MutS 단백질을 고정하였다. MutS 단백질의 고정화 정도를 측정하기 위하여 QCM을 사용하여 공진 주파수의 변화를 측정하여 고정화되는 단백질의 질량변화를 측정하였다. 실험결과 시간의 변화에 따라 진동 주파수는 감소하는 경향을 보이고 있으며 MutS 단백질의 질량변화는 증가하는 경향임을 확인할 수 있다. 그러므로 MutS 단백질이 NTA 표면에 효과적으로 화학결합을 하여 질량의 변화가 증가하기 때문이다. 이와 같은 실험결과를 통하여 Au표면에 NTA-MutS 단백질을 고정함으로 MutS 단백질이mismatched DNA을 감지할 수 있을 것이라 판단된다.
Cardioprotective Effect of the SDF-1α/CXCR4 Axis in Ischemic Postconditioning in Isolated Rat Hearts
김정수,장영호,김준홍,박용현,황선애,김준,이성률,Zhelong Xu,반창일,안교한,전국진 대한심장학회 2017 Korean Circulation Journal Vol.47 No.6
Background and Objectives: Information about the role of the stromal cell-derived factor-1α (SDF-1α)/chemokine receptor type 4 (CXCR4) axis in ischemic postconditioning (IPOC) is currently limited. We hypothesized that the SDF-1α/CXCR4 signaling pathway is directly involved in the cardioprotective effect of IPOC. Methods: Isolated rat hearts were divided into four groups. The control group was subjected to 30-min of regional ischemia and 2-hour of reperfusion (n=12). The IPOC group was induced with 6 cycles of 10-second reperfusion and 10-second global ischemia (n=8) in each cycle. The CXCR4 antagonist, AMD3100, was applied before reperfusion in the IPOC group (AMD+IPOC group, n=11) and control group (AMD group, n=9). Hemodynamic changes with electrocardiography were monitored and infarct size was measured. The SDF-1α, lactate dehydrogenase (LDH) and creatine kinase (CK) concentrations in perfusate were measured. We also analyzed extracellular signal-regulated kinase 1/2 (ERK1/2) and Akt phosphorylation state expression. Results: IPOC significantly reduced infarct size, but AMD3100 attenuated the infarct reducing effect of IPOC. IPOC significantly decreased LDH and CK, but these effects were reversed by AMD3100. ERK1/2 and Akt phosphorylation increased with IPOC and these effects were blocked by AMD3100. Conclusion: Based on the results of this study, SDF-1α/CXCR4 signaling may be involved in IPOC cardioprotection and this signaling pathway couples to the ERK1/2 and Akt pathways.
Simple Screening Method for Double-strand DNA Binders Using Hairpin DNA-modified Magnetic Beads
Hunho Jo,Kyoungin Min,Kyung-mi Song,구자강,한민수,반창일 대한화학회 2011 Bulletin of the Korean Chemical Society Vol.32 No.1
We designed an effective screening method for double strand DNA (dsDNA) binders using DNA-modified magnetic particles. Hairpin DNA was immobilized on the surface of magnetic particle for a simple screening of dsDNA binding materials in a solution containing various compounds. Through several magnetic separation and incubation processes,four DNA-binding materials, DAPI, 9AA, AQ2A, and DNR, were successfully screened from among five candidates. Efficiency of screening was demonstrated by HPLC analysis using a C2/18 reverse-phase column. In addition, their relative binding strengths were verified by measuring the melting temperature (T_m). If hairpin DNA sequence is modified for other uses, this magnetic bead-based approach can be applied as a high-throughput screening method for various functional materials such as anti-cancer drugs.