• 대한의용생체공학회:의공학회지
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• 제42권3호
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• pp.125-142
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• 2021

The POCT (point-of-care test) sensing that has been a fast-developing field is expected to be a next generation technology in health care. The POCT sensors for the detection of proteins, small molecules and especially nucleic acids have lately attracted considerable attention. According to the World Health Organization (WHO), the POCT methods are required to follow the ASSURED guidelines (Affordable, Sensitive, Specific, User- friendly, Robust and rapid, Equipment-free, Deliverable to all people who need the test). Recently, several CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) based diagnostic techniques using the sensitive gene recognition function of CRISPR have been reported. CRISPR/Cas (Cas, CRISPR associated protein) systems based detection technology is the most innovative gene analysis technology that is following the ASSURED guidelines. It is being re-emerged as a powerful diagnostic tool that can detect nucleic acids due to its characteristics that enable rapid, sensitive and specific analyses of nucleic acid. The first CRISPR-based diagnosis begins with the discovery of the additional function of Cas13a. The enzymatic cleavage occurs when the conjugate of Cas protein and CRISPR RNA (crRNA) detect a specific complementary sequence of the target sequence. Enzymatic cleavage occurs on not only the target sequence, but also all surrounding non-target single-stranded RNAs. This discovery was immediately utilized as a biosensor, and numerous sensor studies using CRISPR have been reported since then. In this review, the concept of CRISPR, the characteristics of the Cas protein required for CRISPR diagnosis, the current research trends of CRISPR diagnostic technology, and some aspects to be improved in the future are covered.

• KSBB Journal
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• 제20권2호
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• pp.93-99
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• 2005

불포화지방산의 생합성능이 뛰어나다고 알려진 Thraustochytrids 균주 중 26185를 대상으로 arachidonic acid (AA C20:4 n-6)를 포함하는 PUFA 생산에 관여하는 유전자의 하나로서 key enzyme인 desaturase를 보다 효율적이고 안정적인 방법으로 cloning한 후 결과물을 분석하였다. 이를 위해 실험균주인 26185 균주를 대상으로 효율적인 전체 nucleic acids 추출법을 개발하였으며 관련된 기법의 활용을 통해 일반적으로 활용이 가능한 효과적이 방법임을 확인하였다. 확보된 유전체를 통한 direct PCR의 결과물을 기존에 알려진 cDNA 합성방법에 의한 결과물과 비교하였을 때 동일한 결과물임을 확인하였다. 이는 Thraustochytrids 관련 균주로부터 지방산 생합성에 관련된 효소군을 탐색하는 방법이 cDNA 합성방법에 의하지 않고 보다 직접적으로 진행될 수 있음을 제안할 수 있는 하나의 결과물로 생각되어진다. 획득된 유전자를 같은 진핵세포인 P. pastoris를 숙주로 하여 유전자를 도입하고 활성을 재현성 있게 규명함으로써 인공진화 혹은 재조합균체의 개발이 가능하다는 사실을 부분적으로 제시할 수 있었다.

• Tuberculosis and Respiratory Diseases
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• 제74권6호
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• pp.251-255
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• 2013

Tuberculosis (TB) is one of the largest health problems in the world today. And the incidence of nontuberculous mycobacteria (NTM) lung disease appears to be increasing worldwide. Recently, an automated, nucleic acid amplification assay for the rapid detection of both Mycobacterium tuberculosis and rifampin resistance was developed (Xpert MTB/RIF). And fixed-dose combinations of anti-TB drugs and linezolid have been introduced in the treatment of TB. And new NTM species, named Mycobacterium massiliense, which is very closely related to Mycobacterium abscessus was reported. In this review, these recent advances in the diagnosis and treatment of TB and clinical characteristics of M. massiliense lung disease are discussed.

• JSTS:Journal of Semiconductor Technology and Science
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• 제1권4호
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• pp.232-238
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• 2001

A new DNA purification chip is proposed and fabricated for the sample preparation of Nucleic Acid (NA) probe assay. The proposed DNA purification chip is fabricated using photosensitive glass substrate and polydimethylsiloxane (PDMS) cover fixture. We have successfully captured and eluted the DNA using the fabricated photosensitive glass chip. The fabricated DNA purification chip showed a binding capacity of $15ng/\textrm{cm}^2$and a minimum extractable input concentration of $100copies/200\muL$. The proposed DNA purification chip can be applied for low-cost, disposable sample preparation of NA probe assays.

• Journal of Microbiology and Biotechnology
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• 제20권2호
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• pp.287-293
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• 2010

Reliable discrimination of a single nucleotide mismatch was demonstrated using arrays with peptide nucleic acid (PNA) probes. The newly developed PNA probes immobilization method and hybridization conditions for PNA arrays gave excellent specificity and sensitivity. In addition we compared the specificity, sensitivity, and stability obtained with the PNA and DNA arrays in discriminating single nucleotide mismatches. The PNA arrays had superior perfect match-to-mismatch signal ratios and sensitivities. The relative signal intensities of mismatch PNA probes ranged from 1.6% to 12.1% of the perfect-match PNA probes. These results demonstrated that the PNA arrays were 2.0 to 37.3 times more specific and about 10 times more sensitive than DNA arrays. The PNA array showed the same specificity and sensitivity after 12-month storage at room temperature.

• Archives of Pharmacal Research
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• 제26권3호
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• pp.183-191
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• 2003

Nucleic acid therapies represent a direct genetic approach for cancer treatment. Such an approach takes advantage of mechanisms that activate genes known to confer a growth advantage to neoplastic cells. The ability to block the expression of these genes allows exploration of normal growth regulation. Progress in antisense technology has been rapid, and the traditional antisense inhibition of gene expression is now viewed on a genomic scale. This global view has led to a new vision in antisense technology, the elimination of nonspecific and undesirable side effects, and ultimately, the generation of more effective and less toxic nucleic acid medicines. Several antisense oligonucleotides are in clinical trials, are well tolerated, and are potentially active therapeutically. Antisense oligonucleotides are promising molecular medicines for treating human cancer in the near future.

• Bulletin of the Korean Chemical Society
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• 제26권4호
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• pp.629-633
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• 2005

In the reversed-phase chromatography, the retention time of sample was investigated based on the molecular structure of compound. Several descriptors that were related to retention factors were selected, and then the values of descriptors were calculated with several softwares. The effect of retention factor was measured with calculated values, and the results were obtained that each descriptors of molecular structure of compound have different effect on the retention factor. Therefore, the empirical equation for seven types of descriptors considered was obtained, and it has high values of correlation coefficient. Furthermore, the experimental data and calculated values have good agreement.

• Biomolecules & Therapeutics
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• 제21권6호
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• pp.423-434
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• 2013

The adoption of oligonucleotide aptamer is well on the rise, serving an ever increasing demand for versatility in biomedical field. Through the SELEX (Systematic Evolution of Ligands by EXponential enrichment), aptamer that can bind to specific target with high affinity and specificity can be obtained. Aptamers are single-stranded nucleic acid molecules that can fold into complex three-dimensional structures, forming binding pockets and clefts for the specific recognition and tight binding of any given molecular target. Recently, aptamers have attracted much attention because they not only have all of the advantages of antibodies, but also have unique merits such as thermal stability, ease of synthesis, reversibility, and little immunogenicity. The advent of novel technologies is revolutionizing aptamer applications. Aptamers can be easily modified by various chemical reactions to introduce functional groups and/or nucleotide extensions. They can also be conjugated to therapeutic molecules such as drugs, drug containing carriers, toxins, or photosensitizers. Here, we discuss new SELEX strategies and stabilization methods as well as applications in drug delivery and molecular imaging.

• BMB Reports
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• 제43권1호
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• pp.1-8
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• 2010

The cold shock domain (CSD) is among the most ancient and well conserved nucleic acid binding domains from bacteria to higher animals and plants. The CSD facilitates binding to RNA, ssDNA and dsDNA and most functions attributed to cold shock domain proteins are mediated by this nucleic acid binding activity. In prokaryotes, cold shock domain proteins only contain a single CSD and are termed cold shock proteins (Csps). In animal model systems, various auxiliary domains are present in addition to the CSD and are commonly named Y-box proteins. Similar to animal CSPs, plant CSPs contain auxiliary C-terminal domains in addition to their N-terminal CSD. Cold shock domain proteins have been shown to play important roles in development and stress adaptation in wide variety of organisms. In this review, the structure, function and regulation of plant CSPs are compared and contrasted to the characteristics of bacterial and animal CSPs.

• Fisheries and Aquatic Sciences
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• 제17권3호
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• pp.339-344
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• 2014

PCR amplification with universal primer is a useful tool for speciation of symbionts in marine eukaryote coupled with robust separation method such as denaturing high performance chromatography (DHPLC). To overcome the biased amplification, clamping PCR is recommended to suppress the amplification of host gene. In this study, we evaluated the efficiency of rare gene detection for two kinds of clamping probes which were successfully utilized for eukaryotic symbiont analysis: C3 linked nucleotide (C3) and peptide nucleic acid (PNA). PNA was 3-4 orders of magnitude higher than that of C3 tested in clamping efficiency and rare gene detection. This represented that PNA could be a more competent clamping probe for the enhancement of PCR amplification for rare symbiont genes.