• Title/Summary/Keyword: Gene editing

Search Result 112, Processing Time 0.03 seconds

RNA Editing Enzyme ADAR1 Suppresses the Mobility of Cancer Cells via ARPIN

  • Min Ji Park;Eunji Jeong;Eun Ji Lee;Hyeon Ji Choi;Bo Hyun Moon;Keunsoo Kang;Suhwan Chang
    • Molecules and Cells
    • /
    • v.46 no.6
    • /
    • pp.351-359
    • /
    • 2023
  • Deamination of adenine or cytosine in RNA, called RNA editing, is a constitutively active and common modification. The primary role of RNA editing is tagging RNA right after its synthesis so that the endogenous RNA is recognized as self and distinguished from exogenous RNA, such as viral RNA. In addition to this primary function, the direct or indirect effects on gene expression can be utilized in cancer where a high level of RNA editing activity persists. This report identified actin-related protein 2/3 complex inhibitor (ARPIN) as a target of ADAR1 in breast cancer cells. Our comparative RNA sequencing analysis in MCF7 cells revealed that the expression of ARPIN was decreased upon ADAR1 depletion with altered editing on its 3'UTR. However, the expression changes of ARPIN were not dependent on 3'UTR editing but relied on three microRNAs acting on ARPIN. As a result, we found that the migration and invasion of cancer cells were profoundly increased by ADAR1 depletion, and this cellular phenotype was reversed by the exogenous ARPIN expression. Altogether, our data suggest that ADAR1 suppresses breast cancer cell mobility via the upregulation of ARPIN.

CRISPR and Target-Specific DNA Endonucleases for Efficient DNA Knock-in in Eukaryotic Genomes

  • Lee, Seung Hwan;Kim, Sunghyun;Hur, Junho K
    • Molecules and Cells
    • /
    • v.41 no.11
    • /
    • pp.943-952
    • /
    • 2018
  • The discovery and mechanistic understanding of target-specific genome engineering technologies has led to extremely effective and specific genome editing in higher organisms. Target-specific genetic modification technology is expected to have a leading position in future gene therapy development, and has a ripple effect on various basic and applied studies. However, several problems remain and hinder efficient and specific editing of target genomic loci. The issues are particularly critical in precise targeted insertion of external DNA sequences into genomes. Here, we discuss some recent efforts to overcome such problems and present a perspective of future genome editing technologies.

DNA Barcoding for the Hydrothermal Vent Crab Austinograea Species (Crustacea: Bythograeidae) from the North Fiji Basin, Southwestern Pacific Ocean

  • Lee, Won-Kyung;Ju, Se-Jong;Hou, Bo Kyeng;Kim, Se-Joo
    • Animal Systematics, Evolution and Diversity
    • /
    • v.35 no.1
    • /
    • pp.30-32
    • /
    • 2019
  • The brachyuran crab Bythograeidae Williams, 1980 is common in hydrothermal vent fields worldwide and has recorded to sixteen species of six genera. In this study, we firstly determined the cytochrome c oxidase subunit 1 (CO1) DNA barcodes for the fifth species of Austinograea, A. hourdezi, from hydrothermal vent regions of the North Fiji Basin in southwestern Pacific Ocean. All CO1 DNA barcodes of A. hourdezi were identical. The interspecies variations of three bythograeid genera were 10.9-13.3% for Austinograea, 6.6-15.7% for Bythograea, and 9.7% for Gandalfus. These results would be helpful to understand taxonomy of brachyuran crabs living in hydrothermal vent fields using CO1 DNA barcodes.

Generation of knockout mouse models of cyclin-dependent kinase inhibitors by engineered nuclease-mediated genome editing

  • Park, Bo Min;Roh, Jae-il;Lee, Jaehoon;Lee, Han-Woong
    • Laboraroty Animal Research
    • /
    • v.34 no.4
    • /
    • pp.264-269
    • /
    • 2018
  • Cell cycle dysfunction can cause severe diseases, including neurodegenerative disease and cancer. Mutations in cyclin-dependent kinase inhibitors controlling the G1 phase of the cell cycle are prevalent in various cancers. Mice lacking the tumor suppressors $p16^{Ink4a}$ (Cdkn2a, cyclin-dependent kinase inhibitor 2a), $p19^{Arf}$ (an alternative reading frame product of Cdkn2a,), and $p27^{Kip1}$ (Cdkn1b, cyclin-dependent kinase inhibitor 1b) result in malignant progression of epithelial cancers, sarcomas, and melanomas, respectively. Here, we generated knockout mouse models for each of these three cyclin-dependent kinase inhibitors using engineered nucleases. The $p16^{Ink4a}$ and $p19^{Arf}$ knockout mice were generated via transcription activator-like effector nucleases (TALENs), and $p27^{Kip1}$ knockout mice via clustered regularly interspaced short palindromic repeats/CRISPR-associated nuclease 9 (CRISPR/Cas9). These gene editing technologies were targeted to the first exon of each gene, to induce frameshifts producing premature termination codons. Unlike preexisting embryonic stem cell-based knockout mice, our mouse models are free from selectable markers or other external gene insertions, permitting more precise study of cell cycle-related diseases without confounding influences of foreign DNA.

CDRgator: An Integrative Navigator of Cancer Drug Resistance Gene Signatures

  • Jang, Su-Kyeong;Yoon, Byung-Ha;Kang, Seung Min;Yoon, Yeo-Gha;Kim, Seon-Young;Kim, Wankyu
    • Molecules and Cells
    • /
    • v.42 no.3
    • /
    • pp.237-244
    • /
    • 2019
  • Understanding the mechanisms of cancer drug resistance is a critical challenge in cancer therapy. For many cancer drugs, various resistance mechanisms have been identified such as target alteration, alternative signaling pathways, epithelial-mesenchymal transition, and epigenetic modulation. Resistance may arise via multiple mechanisms even for a single drug, making it necessary to investigate multiple independent models for comprehensive understanding and therapeutic application. In particular, we hypothesize that different resistance processes result in distinct gene expression changes. Here, we present a web-based database, CDRgator (Cancer Drug Resistance navigator) for comparative analysis of gene expression signatures of cancer drug resistance. Resistance signatures were extracted from two different types of datasets. First, resistance signatures were extracted from transcriptomic profiles of cancer cells or patient samples and their resistance-induced counterparts for >30 cancer drugs. Second, drug resistance group signatures were also extracted from two large-scale drug sensitivity datasets representing ~1,000 cancer cell lines. All the datasets are available for download, and are conveniently accessible based on drug class and cancer type, along with analytic features such as clustering analysis, multidimensional scaling, and pathway analysis. CDRgator allows meta-analysis of independent resistance models for more comprehensive understanding of drug-resistance mechanisms that is difficult to accomplish with individual datasets alone (database URL: http://cdrgator.ewha.ac.kr).

Strengthening the competitiveness of agricultural biotechnology through practical application of gene editing technology (유전자편집 작물의 개발 현황 및 농업생명공학기술의 국가 경쟁력 강화)

  • Lee, Shin-Woo
    • Journal of Plant Biotechnology
    • /
    • v.45 no.3
    • /
    • pp.155-170
    • /
    • 2018
  • In this paper, mechanisms of gene editing technologies including ZFN, TALENS and CRISPR were briefly discussed with mutual advantages and disadvantages. Classification criteria of gene edited, site-directed mutagenesis (SDN) crops for regulatory purpose were also discussed. The number of studies using CRISPR technology was high and studies conducted on Arabidopsis thaliana and rice were highest, followed by tobacco, tomato, wheat, and corn. It has been applied to a variety of plants such as other grain crops, flower crops, vegetable crops, and fruit trees. The number of studies focused on practical application or commercialization in the future were also increasing yearly, and the scope of studies also expanded to include research on metabolic engineering for mass production of useful proteins or substances, development of disease resistant crops against viruses, bacteria, and fungi, abiotic environmental stressresistant crops, and increased yields. In addition to this, it was revealed that application range is becoming more diversified, including the development of parthenocarpic tomatoes, hybrid rice lines using male sterility and increased shattering resistance Brassica napus. It was also revealed that the number of CRISPR gene edited crops permitted by the USDA(APHIS) increases yearly, to be released in the international seed market soon.

Determination of the Length of Target Recognition Sequence in sgRNA Required for CRISPR Interference (CRISPR 간섭에 필요한 sgRNA 표적 인식 서열 길이의 결정)

  • Kim, Bumjoon;Kim, Byeong Chan;Lee, Ho Joung;Lee, Sang Jun
    • Microbiology and Biotechnology Letters
    • /
    • v.49 no.4
    • /
    • pp.534-542
    • /
    • 2021
  • Single-molecular guide RNA (sgRNA) plays a role in recognizing the DNA target sequence in CRISPR technology for genome editing and gene expression control. In this study, we systematically compared the length of the target recognition sequence in sgRNAs required for genome editing using Cas9-NG (an engineered Cas9 recognizing 5'-NG as PAM sequence) and gene expression control using deactivated Cas9-NG (dCas9-NG) by targeting the gal promoter in E. coli. In the case of genome editing, the truncation of three nucleotides in the target recognition sequence (TRS) of sgRNA was allowed. In gene expression regulation, we observed that target recognition and binding were possible even if eleven nucleotides were deleted from twenty nucleotides of the TRS. When 4 or more nucleotides are truncated in the TRS of the sgRNA, it is thought that the sgRNA/Cas9-NG complex can specifically bind to the target DNA sequence, but lacks endonuclease activity to perform genome editing. Our study will be helpful in the development of artificial transcription factors and various CRISPR technologies in the field of synthetic biology.