• 제목/요약/키워드: gene targeting

검색결과 483건 처리시간 0.035초

Cadmium chloride down-regulates the expression of Rad51 in HC11 cells and reduces knock-in efficiency

  • Ga-Yeon Kim;Man-Jong Kang
    • 한국동물생명공학회지
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    • 제38권3호
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    • pp.99-108
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    • 2023
  • Background: Efficient gene editing technology is needed for successful knock-in. Homologous recombination (HR) is a major double-strand break repair pathway that can be utilized for accurately inserting foreign genes into the genome. HR occurs during the S/G2 phase, and the DNA mismatch repair (MMR) pathway is inextricably linked to HR to maintain HR fidelity. This study was conducted to investigate the effect of inhibiting MMR-related genes using CdCl2, an MMR-related gene inhibitor, on HR efficiency in HC11 cells. Methods: The mRNA and protein expression levels of MMR-related genes (Msh2, Msh3, Msh6, Mlh1, Pms2), the HR-related gene Rad51, and the NHEJ-related gene DNA Ligase IV were assessed in HC11 cells treated with 10 μM of CdCl2 for 48 hours. In addition, HC11 cells were transfected with a CRISPR/sgRNA expression vector and a knock-in vector targeting Exon3 of the mouse-beta casein locus, and treated with 10 μM cadmium for 48 hours. The knock-in efficiency was monitored through PCR. Results: The treatment of HC11 cells with a high-dose of CdCl2 decreased the mRNA expression of the HR-related gene Rad51 in HC11 cells. In addition, the inhibition of MMR-related genes through CdCl2 treatment did not lead to an increase in knock-in efficiency. Conclusions: The inhibition of MMR-related gene expression through high-dose CdCl2 treatment reduces the expression of the HR-related gene Rad51, which is active during recombination. Therefore, it was determined that CdCl2 is an inappropriate compound for improving HR efficiency.

In Vivo Roles of Lysophospholipid Receptors Revealed by Gene Targeting Studies in Mice

  • Ishii, Isao
    • 대한약학회:학술대회논문집
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    • 대한약학회 2002년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2
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    • pp.96-97
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    • 2002
  • Lysophosphatidic acid (LPA) and sphingosine 1-phosphate (SIP) are bioactive lysophospholipids (LPs) that act as mediators in various cellular processes, such as cell growth, differentiation, survival, motility, and cytoskeletal reorganization (1,2). LPA and S1P are both abundant in serum and are produced by activated platelets and other cell types. (omitted)

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Genetic Suppressor Elements that Halt the Proliferation of Breast Carcinoma Cells

  • Primiano, Thomas
    • 대한약학회:학술대회논문집
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    • 대한약학회 2002년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2
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    • pp.98-114
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    • 2002
  • The completion of the draft sequence of the human genome has provided us with a partial list of known and putative human genes, the total number of which is estimated between 30, 000 and 45, 000 (1, 2). These genes provide many potential targets for drugs, some of which may be useful in stopping the growth of cancers. The development of gene-targeting anticancer drugs could be greatly facilitated by the ability to narrow down the list of human genes to those that are necessary for the growth of tumor cells. (omitted)

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Overview of ALK and ROS1 Rearranged Lung Cancer

  • Choi, Chang Min
    • Tuberculosis and Respiratory Diseases
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    • 제75권6호
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    • pp.236-237
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    • 2013
  • Many attempts have been made to find genetic abnormalities inducing carcinogenesis after the development of epidermal growth factor receptor (EGFR)-tyrosine kinase inhibitor targeting EGFR in lung cancer. New target therapies have been already commercialized and studied along with the recent discovery of gene rearrangement involved in the carcinogenic process of non-small cell lung cancer. This study aims to investigate anplastic lymphoma kinase, c-ros oncogene 1, and receptor tyrosine kinase, in particular.

GFP as a Genetic Marker in Transgenic Fish

  • Lee, Jeong-Ho;Kim, Kyung-Kil;Kim, Young-Ok
    • 한국어업기술학회:학술대회논문집
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    • 한국어업기술학회 2002년도 추계 수산관련학회 공동학술대회발표요지집
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    • pp.183-184
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    • 2002
  • The use of transgenic fish has so far been chiefly limited by the lack of predictable, strong, tissue specific, and position-independent expression of transgenes. For genetic analysis, expression of a marker transgene, easily screenable in the living fish, could facilitate studies of gene targeting, insertional mutagenesis, lineage, and mutational analysis. (omitted)

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Prevalence of Feline Hemotropic Mycoplasmas Among Feral Cats in Korea by use of a PCR assay

  • Cho, Eun Kwang;Yu, DoHyeon;Choi, Ul Soo
    • 한국임상수의학회지
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    • 제33권3호
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    • pp.145-150
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    • 2016
  • We determined the prevalence of feline hemotropic mycoplasma species including 'Candidatus Mycoplasma haemominutum', Mycoplasma haemofelis, and 'Candidatus Mycoplasma turicensis' in naturally infected feral cats in Jeonju, Korea. Forty six feral cats were evaluated by PCR assay targeting the 16S rRNA gene sequence. Nine cats (19.6%) were positive for 'Candidatus Mycoplasma haemominutum', 2 cats (4.3%) were positive for 'Mycoplasm a haemofelis', and 1 cat (2.2%) was infected with both 'Candidatus Mycoplasma haemominutum' and Mycoplasma haemofelis. 'Candidatus Mycoplasma turicensis' was undetected. Partial 16S rRNA gene sequences of Mycoplasma haemofelis were closely (> 96%) related to those from other countries. The amplification of hemoplasma DNA in these samples confirmed the presence of 'Candidatus M. haemominutum' and M. haemofelis in Korea.

Identification of Genes for Mycothiol Biosynthesis in Streptomyces coelicolor A3(2)

  • Park Joo-Hong;Cha Chang-Jun;Roe Jung-Hye
    • Journal of Microbiology
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    • 제44권1호
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    • pp.121-125
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    • 2006
  • Mycothiol is a low molecular weight thiol compound produced by a number of actinomycetes, and has been suggested to serve both anti-oxidative and detoxifying roles. To investigate the metabolism and the role of mycothiol in Streptomyces coelicolor, the biosynthetic genes (mshA, B, C, and D) were predicted based on sequence homology with the mycobacterial genes and confirmed experimentally. Disruption of the mshA, C, and D genes by PCR targeting mutagenesis resulted in no synthesis of mycothiol, whereas the mshB mutation reduced its level to about $10\%$ of the wild type. The results indicate that the mshA, C, and D genes encode non-redundant biosynthetic enzymes, whereas the enzymatic activity of MshB (acetylase) is shared by at least one other gene product, most likely the mca gene product (amidase).

A Novel Marker for the Species-Specific Detection and Quantitation of Shigella sonnei by Targeting a Methylase Gene

  • Cho, Min Seok;Ahn, Tae-Young;Joh, Kiseong;Kwon, Oh-Sang;Jheong, Won-Hwa;Park, Dong Suk
    • Journal of Microbiology and Biotechnology
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    • 제22권8호
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    • pp.1113-1117
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    • 2012
  • Shigella sonnei is a causal agent of fever, nausea, stomach cramps, vomiting, and diarrheal disease. The present study describes a quantitative polymerase chain reaction (qPCR) assay for the specific detection of S. sonnei using a primer pair based on the methylase gene for the amplification of a 325 bp DNA fragment. The qPCR primer set for the accurate diagnosis of Shigella sonnei was developed from publically available genome sequences. This quantitative PCR-based method will potentially simplify and facilitate the diagnosis of this pathogen and guide disease management.