• 제목/요약/키워드: gene replacement

검색결과 134건 처리시간 0.022초

Selection of L-arabinase gene to degrade Corn fiber

  • Ahn, Mi-Sun;Lee, Hyoung-Joo;Ryu, Yeon-Woo
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2005년도 생물공학의 동향(XVI)
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    • pp.317-321
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    • 2005
  • L- arabinose residues are widely distributed in plant cell walls, where they are present in polymers such as arabinans, arabinoxylans, arabinogalactans and arabinogalactan proteins. L-arabinose suppress intestinal sucrase and decrease the adsorption of sugar in the small intestine, consequently, weight loss and fatness prevent. Now, xylose be used replacement sugar and arabinose be utilized fatness prevent of our time. Various Agricultural surplus like com fiber, contain $20\;{\sim}\;40%$ of hemicellulose. Corn fiber from Agricultural Renewable Biomass was chosen the best suitable material for arabinose production. In this work, we searched about for L-arabinose gene in compost, metagenome pool and indonesian soil. So, the B1029 TS2-8 of L-arabinase gene in compost was selected by YNB media(5% yeast nitrogen base, 5% arabinogalactan). After enzyme reaction with corn fiver, B1029 TS2-8 produced 2.15 g/L of L-arabonose.

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Novel Therapeutic Approaches to Mucopolysaccharidosis Type III

  • Yang, Aram
    • Journal of mucopolysaccharidosis and rare diseases
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    • 제5권1호
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    • pp.22-28
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    • 2021
  • Mucopolysaccharidosis type III (MPS III) or Sanfilippo disease is an orphan-inherited lysosomal storage disease. It is one of the most common MPS subtypes. The classical presentation is an infantile-onset neurodegenerative disease characterized by intellectual regression, behavioral and sleep disturbances, loss of ambulation, and early death. Unlike other MPS, no disease-modifying therapy has been approved. Here, we review the curative therapy developed for MPS III, from historically ineffective hematopoietic stem cell transplantation and substrate reduction therapy to the promising enzyme replacement therapy or adeno-associated/lentiviral vector-mediated gene therapy. Preclinical studies are presented with recent translational first-in-man trials. We also present experimental research with preclinical mRNA and gene-editing strategies. Lessons from animal studies and clinical trials have highlighted the importance of early therapy before extensive neuronal loss. Disease-modifying therapy for MPS III will likely mandate the development of new early diagnosis strategies.

Cryparin 유전자의 promoter 분석을 위한 cryparin 유전자 치환체의 순수 제조 (Construction of a Pure Cryparin-null Mutant for the Promoter Analysis of Cryparin Gene)

  • 김명주;양문식;김대혁
    • 한국균학회지
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    • 제26권4호통권87호
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    • pp.450-457
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    • 1998
  • Cryparin은 Cryphonectria parasitica의 세포벽에 풍부한 소수성 단백질에 속한다. cryparin은 비록 하나의 유전자에 의해 발현되지만 액체배양 후 48시간이 지나면 발현된 전체 유전자중에서 22%를 차지할 정도의 높은 발현 양상을 나타낸다. 또한 cryparin은 RNA mycovirus인 Cryphonectria hypovirus 1의 감염에 의해 발현이 현저히 억제되는 유전자로 알려졌다. 이미 지난 실험(Kim et al., 1999)에서 상동염색체간의 재조합을 이용하여 cryparin 유전자를 항생제 hygromycin B 저항성 유전자로 치환한 치환체를 제조하였다. 발현율이 매우 높으면서도 virus에 의해 밀접하게 영향받는 cryparin 유전자의 promoter 분석을 위하여서는 대상이 되는 유전자 치환을 위한 vector만을 포함하며, 분석에 이용될 여러 유전자 운반체들이 어느 한곳에만 삽입되도록 하는 성질을 가진 균주의 개발이 필요하다. 그러나 지난번 실험의 결과 얻어진 cryparin 치환체는 치환용 vector외에도 무작위로 삽입된 vector가 존재하고 나아가 새로운 vector들이 어느 한곳에만 삽입되도록 하는 성질을 갖지 못하였다. 따라서 본 실험에서는 cryparin 유전자 치환체와 영양요구성 돌연변이체인 균주간의 교잡을 이용하여 분석 대상이 되는 유전자의 치환에 이용된 vector만을 포함하며, 분석에 이용될 여러 유전자 운반체들이 genome내의 어느 한곳에만 삽입되도록 하는 성질을 가진 균주를 제조하였다.

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Transferrine peptide ligand로 개량된 아데노바이러스를 이용한 신경전구세포로의 유전자 전달 효율 조사 (Modified Adenovirus Mediated Gene Transfer to Neuronal Precursor Cells)

  • 정인실
    • 미생물학회지
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    • 제42권1호
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    • pp.73-76
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    • 2006
  • 신정전구세포를 이용한 퇴행성 뇌질환의 세포치료나 유전자치료에서 효율적인 유전자 전달을 목적으로 개량된 아데노바이러스 벡터의 실용 가능성을 쥐의 해마에서 유래된 신정전구세포를 이용하여 조사하였다. 외피단백질을 조작한 개략 아데노바이러스벡터는 분화전과 후의 신정전구세포로 1세대 아데노바이러스 벡터에 비해 6배 정도 유전자를 효율적으로 전달하였다. 또한 바이러스의 감염은 신정전구세포가 신경세포나 신정 아교세포로 분화하는데 영향을 미치는 않았다. 따라서 신정전구세포를 이용한 신정질환의 세포치료나 유전자 치료에서 개량된 아데노바이러스로 유전자를 전달하면 치료의 효율성을 향상시킬 수 있을 것이다.

The Rat Myosin Light Chain Promoter-Driven DsRed Reporter System Allows Specific Monitoring of Bone Marrow Mesenchymal Stem Cell- Derived Cardiomyocytes

  • Choi, Seung-Cheol;Lim, Do-Sun
    • Reproductive and Developmental Biology
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    • 제32권1호
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    • pp.21-25
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    • 2008
  • Bone marrow mesenchymal stem cells (BMMSCs) have the capacity for self-renewal and differentiation into a variety of cell types. They represent an attractive source of cells for gene and cell therapy. The purpose of this study is to direct the specific expression of the DsRed reporter gene in $Sca-1^+$ BMMSCs differentiated into a cardiomyogenic lineage. We constructed the prMLC-2v-DsRed vector expressing DsRed under the control of the 309 tp fragment of the rat MLC-2v 5'-flanking region. The specific expression of the DsRed reporter gene under the transcriptional control of the 309 bp fragment of the rat MLC-2v promoter was tested in 5-azacytidine healed-$Sca-1^+$ BMMSCs over 2 weeks after the prMLC-2v-DsRed transfection. The prMLC-2v-DsRed was specifically expressed in the $Sca-1^+$ BMMSCs with cardiomyogenic lineage differentiation and it demonstrates that the 309 bp sequences of the rat MLC-2v 5'-flanking region is sufficient to confer cardiac specific expression on a DsRed reporter gene. The cardiac-specific promoter-driven reporter vector provides an important tool for the study of stem cell differentiation and cell replacement therapy in ischemic cardiomyopathy.

Effects of dietary lipid sources on apoptotic and immune gene expression in head kidney of olive flounder (Paralichthys olivaceus)

  • Hur, Deokhwe;Lee, Sang-Min;Hong, Suhee
    • 한국어병학회지
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    • 제25권3호
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    • pp.199-210
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    • 2012
  • It can be hypothesized that dietary fatty acids can modulate immune responses in fish by inducing apoptosis of immune cells since dietary polyunsaturated fatty acid (PUFA) increase apoptosis by oxygen radicals generated by peroxidation. Thus we examined the effects of deferent dietary lipid sources such as squid liver oil (FO), linseed oil (LO) and soybean oil (SO) on oxidation (Cytochrome C oxidase; COS), apoptosis (TNF-${\alpha}$ Scinderin like) and immune (IL-$1{\beta}$ and NKEF) gene expression in the main immune organ (head kidney) in olive flounder (Paralichthys olivaceus) by Q-PCR analysis after feeding diets containing each oil (5%) for 15 weeks. Linseed oil and soybean oil were chosen to compare n-3 or n-6 enriched vegetable oils, respectively. Consequently, COS, TNF-${\alpha}$ and Scinderin like gene expression was increased in SO group, indicating the induction of oxidation and apoptosis. Meanwhile, no significant difference was found in immune gene expression. In conclusion vegetable oils containing n-3 PUFA like linseed oil seems to be more suitable lipid source than soybean oil for replacement of fish oil in flounder since n-6 PUFA in SO leads to activation of apoptosis pathways within the cellular damage in head kidney.

RNA Mapping of Mutant Myotonic Dystrophy Protein Kinase 3'-Untranslated Region Transcripts

  • Song, Min-Sun;Lee, Seong-Wook
    • Genomics & Informatics
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    • 제7권4호
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    • pp.181-186
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    • 2009
  • Myotonic dystrophy type 1 (DM1), which is a dominantly inherited neurodegenerative disorder, results from a CTG trinucleotide repeat expansion in the 3'-untranslated region (3'-UTR) of the myotonic dystrophy protein kinase (DMPK) gene. Retention of mutant DMPK (mDMPK) transcripts in the nuclei of affected cells has been known to be the main cause of pathogenesis of the disease. Thus, reducing the RNA toxicity through elimination of the mutant RNA has been suggested as one therapeutic strategy against DM1. In this study, we suggested RNA replacement with a trans -splicing ribozyme as an alternate genetic therapeutic approach for amelioration of DM1. To this end, we identified the regions of mDMPK 3'-UTR RNA that were accessible to ribozymes by using an RNA mapping strategy based on a trans-splicing ribozyme library. We found that particularly accessible sites were present not only upstream but also downstream of the expanded repeat sequence. Repair or replacement of the mDMPK transcript with the specific ribozyme will be useful for DM1 treatment through reduction of toxic mutant transcripts and simultaneously restore wild-type DMPK or release nucleus-entrapped mDMPK transcripts to the cytoplasm.

Genetically Modified Human Embryonic Stem Cells Relieve Symptomatic Motor Behavior in a Rat Model of Parkinson′s Disease

  • 길광수;이영재;김은영;이창현;이훈택;정길생;박세필;임진호
    • 한국동물번식학회:학술대회논문집
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    • 한국동물번식학회 2003년도 학술발표대회 발표논문초록집
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    • pp.74-74
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    • 2003
  • Embryonic stem cells have several characteristics suitable for cell replacement therapy. To investigate a possibility of using human embryonic stem cell (hESC) as a carrier of therapeutic gene(s), hESC (MB03) was co-transfected with cDNAS coding for tyrosine hydroxylase (TH) and GTP cyclohydrolase Ⅰ (GTPCH Ⅰ) and bulk-selected using neomycin and hygromycin-B. Successful transfection was confirmed by western immunoblotting and RT-PCR. The genetically modified hESC (bk-THGC) relieved apomorphine-induced asymmetric motor behavior by approximately 54% when grafted into striatum of 6-OHDA-denervated rat brain. The number of rotation, however, increased up to 176+18% in 6 weeks when sham-grafted compared with number of rotation before graft. Immunohistochemical staining revealed that the grafted hESC survived and expressed TH for at least 6 weeks while the experiment was continued.

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Site-Specific Mutagenesis of the gshI Gene for Increasing the Activity of ${\gamma}$-Glutamylcysteine Synthetase in Escherichia coli K-12

  • Kwak, Joon-Hyeok;Nam, Yong-Suk;Lee, Se-Yong
    • BMB Reports
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    • 제31권3호
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    • pp.254-257
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    • 1998
  • The gshI gene from the Escherichia coli K-12 strain codes for ${\gamma}-glutamylcysteine$ synthetase which mediates the rate-limiting step of glutathione biosynthesis. The isolated gshI gene from E. coli K-12 has an unusual translation initiation codon, UUG. The 494th amino acid is Ala rather than Gly which was found in a mutant strain E. coli B. In order to improve the translational rate of the gshI gene of E. coli K-12, the initiation codon, UUG, was changed to the usual AUG codon by the site-specific mutagenesis. This change has resulted in a 53% increase of ${\gamma}-glutamylcysteine$ synthetase activity. The enzyme activity was also improved by replacing $Ala^{494}$ with Val (A494V) or Leu (A494L). The replacement of $Ser^{495}$ with Thr (S495T) also resulted in a 62% increase of the enzyme activity. Therefore, the specific activity of ${\gamma}-glutamylcysteine$ synthetase was increased with the increasing chain length of the aliphathic amino acid at the site of the 494th amino acid (Ala<$Val{\leq}Leu$).

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Nomenclature of emerging therapeutics in neurology

  • Shin, Jin-Hong;Park, Young-Eun;Kim, Dae-Seong
    • Annals of Clinical Neurophysiology
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    • 제23권1호
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    • pp.29-34
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    • 2021
  • New therapeutics in neurology are expanding at an unprecedented pace. In addition to the classic enzyme-replacement therapies, monoclonal antibodies are increasingly being used to modulate autoimmunity. RNA therapeutics are an emerging class, together with gene and cell therapies. The nomenclature of international nonproprietary names helps us to recognize these new drugs according to their class and function. Suffixes denote major categories of the drug, while infixes provide additional information such as the source and target.