• Title/Summary/Keyword: Microenviroment

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Stimulation of Oligonucleotide-Directed Gene Correction by Redβ Expression and MSH2 Depletion in Human HT1080 Cells

  • Xu, Ke;Stewart, A. Francis;Porter, Andrew C.G.
    • Molecules and Cells
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    • v.38 no.1
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    • pp.33-39
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    • 2015
  • The correction of disease-causing mutations by single-strand oligonucleotide-templated DNA repair (ssOR) is an attractive approach to gene therapy, but major improvements in ssOR efficiency and consistency are needed. The mechanism of ssOR is poorly understood but may involve annealing of oligonucleotides to transiently exposed single-stranded regions in the target duplex. In bacteria and yeast it has been shown that ssOR is promoted by expression of $Red{\beta}$, a single-strand DNA annealing protein from bacteriophage lambda. Here we show that $Red{\beta}$ expression is well tolerated in a human cell line where it consistently promotes ssOR. By use of short interfering RNA, we also show that ssOR is stimulated by the transient depletion of the endogenous DNA mismatch repair protein MSH2. Furthermore, we find that the effects of $Red{\beta}$ expression and MSH2 depletion on ssOR can be combined with a degree of cooperativity. These results suggest that oligonucleotide annealing and mismatch recognition are distinct but interdependent events in ssOR that can be usefully modulated in gene correction strategies.

The Present and Future of the Cancer Microenvironment Bioprinting (암 미세환경 생체 인쇄의 현재와 미래)

  • Cho, Min Ji;Chi, Byung Hoon;Kim, Myeong Joo;Whang, Young Mi;Chang, In Ho
    • The Korean Journal of Urological Oncology
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    • 제15권3호
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    • pp.103-110
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    • 2017
  • Cancer is the tissue complex consisted with heterogeneous cellular compositions, and microenvironmental cues. During the various stages of cancer initiation, development, and metastasis, cell-cell interactions as well as cell-extracellular matrix play major roles. Conventional cancer models both 2-dimensional and 3-dimensional (3D) present numerous limitations, which restrict their use as biomimetic models for drug screening and fundamental cancer biology studies. Recently, bioprinting biofabrication platform enables the creation of high-resolution 3D structures. Moreover this platform has been extensively used to model multiple organs and diseases, and this versatile technique has further found its creation of accurate models that figure out the complexity of the cancer microenvironment. In this review we will focus on cancer biology and limitations with current cancer models and we discuss vascular structures bioprinting that are critical to the construction of complex 3D cancer organoids. We finally conclude with current literature on bioprinting cancer models and propose future perspectives.

Enzymatic Synthesis of Meth.yl Fructoside by Immobilized Invertase (고정화 전화당 효소에 의한 메틸 프룩토시드의 합성)

  • 허주형;김해성
    • KSBB Journal
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    • v.8 no.4
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    • pp.313-319
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    • 1993
  • Methyl fructoside was synthesized from sucrose and methanol using an immobilized invertase. The enzyme was covalently bound by glutaraldehyde on porous silica coated with polyethyleneimine to give loading capacity of 120mg of invertase per one gram of dry porous silica and effective activity of 100U per one milligram of bound invertase. Polyethyleneimine coating imparted a hydrophillic character, good activity retention and high loading capacity to the surface of porous silica as well as hydrophillic microenviroment in the vicinity of bound invertase. The immobilized enzyme was formed into an alginate-enclosed silica bead to have enough activity for methyl fructoside synthesis from aqueous methanol-sucrose solution. Using the alginate-enclosed biocatalyst the yield of methyl fructoside was obtained as high as 55.9% from aqueous 30% (v/v) methanol and 0.291mo1/l sucrose with 2U/ml activity at $25^{\circ}C$, pH 4.8.

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