• Title/Summary/Keyword: molecular methods

Search Result 3,353, Processing Time 0.032 seconds

Recent advances in developing molecular tools for targeted genome engineering of mammalian cells

  • Lim, Kwang-Il
    • BMB Reports
    • /
    • v.48 no.1
    • /
    • pp.6-12
    • /
    • 2015
  • Various biological molecules naturally existing in diversified species including fungi, bacteria, and bacteriophage have functionalities for DNA binding and processing. The biological molecules have been recently actively engineered for use in customized genome editing of mammalian cells as the molecule-encoding DNA sequence information and the underlying mechanisms how the molecules work are unveiled. Excitingly, multiple novel methods based on the newly constructed artificial molecular tools have enabled modifications of specific endogenous genetic elements in the genome context at efficiencies that are much higher than that of the conventional homologous recombination based methods. This minireview introduces the most recently spotlighted molecular genome engineering tools with their key features and ongoing modifications for better performance. Such ongoing efforts have mainly focused on the removal of the inherent DNA sequence recognition rigidity from the original molecular platforms, the addition of newly tailored targeting functions into the engineered molecules, and the enhancement of their targeting specificity. Effective targeted genome engineering of mammalian cells will enable not only sophisticated genetic studies in the context of the genome, but also widely-applicable universal therapeutics based on the pinpointing and correction of the disease-causing genetic elements within the genome in the near future.

Molecular Application in Psychiatry (정신과의 분자생물학 적용)

  • Choi, Ihn-Geun
    • Korean Journal of Biological Psychiatry
    • /
    • v.7 no.2
    • /
    • pp.115-122
    • /
    • 2000
  • The development of molecular biology has brought many changes in psychiatry. Molecular biology makes us possible to know the cause of mental disorders that provide the way to prevent the disorders, and to develop various accurate diagnostic and treatment methods for mental disorders. The author discusses the concept, cause, and treatment of mental disorders in the aspect of molecular biology. Importing the methods of molecular biology into psychiatry, we can anticipate to get a number of the goals of psychiatric genetics, including identification of specific susceptibility genes, clarification of the pathophysiological processes whereby these genes lead to symptoms, establishment of epigenetic factors that interact with these genes to produce disease, validation of nosological boundaries that more closely reflect the actions of these genes, and development of effective preventive and therapeutic interventions based on genetic counseling, gene therapy, and modification of permissive or protective environmental influences. In addition to their capacity to accelerate the discovery of new molecules participating in the nervous system's response to disease or to self-administered drugs, molecular biological strategies can also be used to determine how critical a particular gene product may be in mediating a cellular event with behavioral importance. Molecular biology probably enables us discover the environmental factors of mental disorders and allow rational drug design and gene therapies for mental disorders, by isolation of gene products that facilitate a basic understanding of the pathogenesis of these disorders. A specific genetic linkage may suggest a novel class of drugs that has not yet been tried. With respect to gene therapy, the hypothetical method would use a gene delivery system, most likely a modified virus, to insert a functional copy of a mutant gene into those brain cells that require the gene for normal function.

  • PDF

Effect of Solubilization Conditions on Molecular Weight Distribution of Enzymatically-Hydrolyzed Silk Peptides (실크의 가용화 조건이 효소분해 실크 펩타이드의 분자량 분포에 미치는 영향)

  • 채희정;인만진;김의용
    • KSBB Journal
    • /
    • v.13 no.1
    • /
    • pp.114-118
    • /
    • 1998
  • The effects of fibron solubilization conditions on molecular weight distribution of enzymatically-hydrolyzed silk peptides were investigated. The weight-averaged molecular weights of silk proteins prepared by solubilization with calcium chloride, ethylenediamine and sulfuric acid were 41600, 3308, and 1268 dalton, respectively. Silk peptides in the average molecular weight range of 600-1200 dalton were obtained by protease treatment from solubilized silk fibroin. After the acid hydrolysis of silk protein using hydrochloric acid for 24 hr, silk protein was hydrolyzed to peptides whose average molecular weight and free amino acid content were 145 dalton and 80%, respectively. It was possible to control molecular weight distribution of silk peptides by the combination of solubilization and hydrolysis methods. Among the various treatment methods, acid solubilization followed by protease treatment had an advantage of molecular weight control for the peptide production.

  • PDF

Molecular prophage typing of Staphylococcus aureus isolates from bovine mastitis

  • Ko, Dae-Sung;Seong, Won-Jin;Kim, Danil;Kim, Eun-Kyung;Kim, Nam-Hyung;Lee, Chung-Young;Kim, Jae-Hong;Kwon, Hyuk-Joon
    • Journal of Veterinary Science
    • /
    • v.19 no.6
    • /
    • pp.771-781
    • /
    • 2018
  • Staphylococcus aureus is one of the major pathogens causing bovine mastitis and foodborne diseases associated with dairy products. To determine the genetic relationships between human and bovine or bovine isolates of S. aureus, various molecular methods have been used. Previously we developed an rpoB sequence typing (RSTing) method for molecular differentiation of S. aureus isolates and identification of RpoB-related antibiotic resistance. In this study, we performed spa typing and RSTing with 84 isolates from mastitic cows (22 farms, 72 cows, and 84 udders) and developed a molecular prophage typing (mPPTing) method for molecular epidemiological analysis of bovine mastitis. To compare the results, human isolates from patients (n = 14) and GenBank (n = 166) were used for real and in silico RSTing and mPPTing, respectively. Based on the results, RST10-2 and RST4-1 were the most common rpoB sequence types (RSTs) in cows and humans, respectively, and most isolates from cows and humans clearly differed. Antibiotic resistance-related RSTs were not detected in the cow isolates. A single dominant prophage type and gradual evolution through prophage acquisition were apparent in most of the tested farms. Thus, RSTing and mPPTing are informative, simple, and economic methods for molecular epidemiological analysis of S. aureus infections.

Effects of Collection Methods on Recovery Efficiency, Maturation Rate and Subsequent Embryonic Developmental Competence of Oocytes in Holstein Cow

  • Wang, Zheng-guang;Yu, Song-dong;Xu, Zi-rong
    • Asian-Australasian Journal of Animal Sciences
    • /
    • v.20 no.4
    • /
    • pp.496-500
    • /
    • 2007
  • Holstein cow ovaries obtained at a slaughterhouse were used to study the influence of the oocyte collection methods (slicing, puncture, aspiration I and II) on recovery efficiency and subsequent in vitro maturation and embryonic development competence of immature oocytes recovered. In the slicing method, the whole ovarian was chopped into small pieces with a surgical blade. In the puncture method, the whole ovarian surface was punctured by 18-g needle. In other 2 aspiration methods, collected oocytes by aspirating from the visible follicles using an 18-g needle attached to a 5 ml syringe (aspiration I) or using a constant negetive pressure (-80 mmHg) with a vacuum pump (aspiration II). The oocytes were classified into 4 classes on the basis of the morphology of cumulus cells and cytoplasmic appearance of oocyte. Slicing ($9.6{\pm}0.4$) and puncture ($9.7{\pm}0.4$)yielded a larger number of oocytes per ovary than other two aspiration methods (aspiration I and II were $5.8{\pm}0.3$and $5.6{\pm}0.4$, respectively) (p<0.05). The number of the highest quality oocytes (grade A) per ovary was significantly higher in slicing ($4.2{\pm}0.2$) and puncture ($4.6{\pm}0.1$) methods than in other methods (aspiration I and II were $1.2{\pm}0.2$ and $1.4{\pm}0.2$, respectively) (p<0.05). The rate of nuclear maturation of the highest and higher quality oocytes (grade A and grade B, respectively) was not affected by the oocytes collection methods. The oocytes collection methods also did not influence subsequent embryonic developmental competence after in vitro fertilization with M II stage oocytes. It is concluded that slicing and puncture methods of the ovaries can be used as an alternative techniques to aspiration by the syringe or vacuum pump.

Molecular Tension Probes to Quantify Cell-Generated Mechanical Forces

  • Baek, Kyung Yup;Kim, Seohyun;Koh, Hye Ran
    • Molecules and Cells
    • /
    • v.45 no.1
    • /
    • pp.26-32
    • /
    • 2022
  • Living cells generate, sense, and respond to mechanical forces through their interaction with neighboring cells or extracellular matrix, thereby regulating diverse cellular processes such as growth, motility, differentiation, and immune responses. Dysregulation of mechanosensitive signaling pathways is found associated with the development and progression of various diseases such as cancer. Yet, little is known about the mechanisms behind mechano-regulation, largely due to the limited availability of tools to study it at the molecular level. The recent development of molecular tension probes allows measurement of cellular forces exerted by single ligand-receptor interaction, which has helped in revealing the hitherto unknown mechanistic details of various mechanosensitive processes in living cells. Here, we provide an introductory overview of two methods based on molecular tension probes, tension gauge tether (TGT), and molecular tension fluorescence microscopy (MTFM). TGT utilizes the irreversible rupture of double-stranded DNA tether upon application of force in the piconewton (pN) range, whereas MTFM utilizes the reversible extension of molecular springs such as polymer or single-stranded DNA hairpin under applied pN forces. Specifically, the underlying principle of how molecular tension probes measure cell-generated mechanical forces and their applications to mechanosensitive biological processes are described.

Standardized microwave extract of Sappan Lignum exerts anti-inflammatory effects through inhibition of NF-κB activation via regulation of heme oxygenase-1 expression

  • Md Anisuzzaman Chowdhury;Moonbum Choi;Wonmin Ko;Hwan Lee;Sam Cheol Kim;Hyuncheol Oh;Eun‑Rhan Woo;Youn‑Chul Kim;Dong‑Sung Lee
    • Molecular Medicine Reports
    • /
    • v.19 no.3
    • /
    • pp.1809-1816
    • /
    • 2019
  • The extract of Sappan Lignum, the heartwood of Caesalpinia sappan L., has been used in medicine to improve blood circulation. Recently, the application of microwave extraction methods has been a major focus of research into the extraction of components from natural sources. In this experiment, we compared the anti-inflammatory effects of Sappan Lignum prepared by heat-70% EtOH extraction (CSE-H-70E) and microwave-70% EtOH extraction (CSE-MW-70E). High-performance liquid chromatography analysis was used to identify the compounds in these extracts. The heat-70% EtOH and microwave-70% EtOH extracts of Sappan Lignum had different chromatograms. CSE-MW-70E significantly inhibited the protein expression of iNOS and COX-2, PGE2, TNF-α, and reduced NO and IL-1β production in macrophages exposed to LPS, whereas, only high concentrations of CSE-H-70E (20 ㎍/ml) resulted in any effects. Furthermore, CSE-MW-70E upregulated heme oxygenase-1 (HO-1) expression. In addition, the use of tin protoporphyrin, an inhibitor of HO-1, confirmed the inhibitory effects of CSE-MW-70E on pro-inflammatory mediators. These results suggested that the CSE-MW-70E-mediated upregulation of HO-1 played an important role in the anti-inflammatory effects of macrophages. Therefore, these findings showed that microwave extraction can be utilized to improve the extraction efficiency and biological activity of Sappan Lignum.

NMR methods in fragment based drug discovery

  • Lim, Jongsoo
    • Journal of the Korean Magnetic Resonance Society
    • /
    • v.19 no.3
    • /
    • pp.132-136
    • /
    • 2015
  • Nuclear magnetic resonance (NMR) spectroscopy, owing to its ability to provide atomic level information on molecular structure, dynamics and interaction, has become one of the most powerful methods in early drug discovery where hit finding and hit-to-lead generation are mainly pursued. In recent years, drug discovery programs originating from the fragment-based drug discovery (FBDD) strategies have been widely incorporated into academia and industry in which a wide variety of NMR methods become an indispensable arsenal to elucidate the binding of small molecules onto bimolecular targets. In this review, I briefly describe FBDD and introduce NMR methods mainly used in FBDD campaigns of my company. In addition, quality control of fragment library and practical NMR methods in industrial aspect are discussed shortly.

Recent Advancement of the Molecular Diagnosis in Pediatric Brain Tumor

  • Bae, Jeong-Mo;Won, Jae-Kyung;Park, Sung-Hye
    • Journal of Korean Neurosurgical Society
    • /
    • v.61 no.3
    • /
    • pp.376-385
    • /
    • 2018
  • Recent discoveries of brain tumor-related genes and fast advances in genomic testing technologies have led to the era of molecular diagnosis of brain tumor. Molecular profiling of brain tumor became the significant step in the diagnosis, the prediction of prognosis and the treatment of brain tumor. Because traditional molecular testing methods have limitations in time and cost for multiple gene tests, next-generation sequencing technologies are rapidly introduced into clinical practice. Targeted sequencing panels using these technologies have been developed for brain tumors. In this article, focused on pediatric brain tumor, key discoveries of brain tumor-related genes are reviewed and cancer panels used in the molecular profiling of brain tumor are discussed.

Molecular Dynamics Simulation of Droplet Vaporization (분자 동력학을 이용한 액적 기화 시뮬레이션)

  • Nam, Gun-Woo;Yoon, Woong-Sup
    • Proceedings of the KSME Conference
    • /
    • 2003.11a
    • /
    • pp.121-126
    • /
    • 2003
  • A study of argon droplet vaporization is conducted using molecular dynamics, instead of using traditional methods such as the Navier-Stokes equation. Molecular dynamics uses Lagrangian frame to describe molecular behavior in a system and uses only momentum and position data of all molecules in the system. So every property is not a hypothetical input but a statistical result calculated from the momentum and position data. This work performed a simulation of the complete vaporization of a three dimensional submicron argon droplet within quiescent environment. Lennard-Jones 12-6 potential function is used as a intermolecular potential function. The molecular configuration is examined while an initially non-spherical droplet is changed into the spherical shape and droplet evaporates. And the droplet radius versus time is calculated with temperature and pressure profile.

  • PDF