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non-homologous end-joining

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https://www.readbyqxmd.com/read/29625543/mechanisms-of-mitochondrial-dna-repair-in-mammals
#1
REVIEW
L A Zinovkina
Accumulation of mutations in mitochondrial DNA leads to the development of severe, currently untreatable diseases. The contribution of these mutations to aging and progress of neurodegenerative diseases is actively studied. Elucidation of DNA repair mechanisms in mitochondria is necessary for both developing approaches to the therapy of diseases caused by mitochondrial mutations and understanding specific features of mitochondrial genome functioning. Mitochondrial DNA repair systems have become a subject of extensive studies only in the last decade due to development of molecular biology methods...
March 2018: Biochemistry. Biokhimii︠a︡
https://www.readbyqxmd.com/read/29622782/multiple-sgrnas-with-overlapping-sequences-enhance-crispr-cas9-mediated-knock-in-efficiency
#2
Da Eun Jang, Jae Young Lee, Jae Hoon Lee, Ok Jae Koo, Hee Sook Bae, Min Hee Jung, Ji Hyun Bae, Woo Sung Hwang, Yoo Jin Chang, Yoon Hoo Lee, Han Woong Lee, Su Cheong Yeom
The CRISPR/Cas9 system is widely applied in genome engineering due to its simplicity and versatility. Although this has revolutionized genome-editing technology, knockin animal generation via homology directed repair (HDR) is not as efficient as nonhomologous end-joining DNA-repair-dependent knockout. Although its double-strand break activity may vary, Cas9 derived from Streptococcus pyogenens allows robust design of single-guide RNAs (sgRNAs) within the target sequence; However, prescreening for different sgRNA activities delays the process of transgenic animal generation...
April 6, 2018: Experimental & Molecular Medicine
https://www.readbyqxmd.com/read/29594212/cell-line-dependent-effects-of-hypoxia-prior-to-irradiation-in-squamous-cell-carcinoma-lines
#3
Franziska Hauth, Mahmoud Toulany, Daniel Zips, Apostolos Menegakis
Purpose: To assess the impact of hypoxia exposure on cellular radiation sensitivity and survival of tumor cells with diverse intrinsic radiation sensitivity under normoxic conditions. Materials and methods: Three squamous cell carcinoma (SCC) cell lines, with pronounced differences in radiation sensitivity, were exposed to hypoxia prior, during or post irradiation. Cells were seeded in parallel for colony formation assay (CFA) and stained for γH2AX foci or processed for western blot analysis...
August 2017: Clinical and Translational Radiation Oncology
https://www.readbyqxmd.com/read/29588483/genome-plasticity-is-governed-by-double-strand-break-dna-repair-in-streptomyces
#4
Grégory Hoff, Claire Bertrand, Emilie Piotrowski, Annabelle Thibessard, Pierre Leblond
The linear chromosome of the bacterium Streptomyces exhibits a remarkable genetic organization with grossly a central conserved region flanked by variable chromosomal arms. The terminal diversity co-locates with an intense DNA plasticity including the occurrence of large deletions associated to circularization and chromosomal arm exchange. These observations prompted us to assess the role of double strand break (DSB) repair in chromosome plasticity following. For that purpose, DSBs were induced along the chromosome using the meganuclease I-SceI...
March 27, 2018: Scientific Reports
https://www.readbyqxmd.com/read/29582426/prevalence-of-brca1-2-large-genomic-rearrangements-in-chinese-women-with-sporadic-triple-negative-or-familial-breast-cancer
#5
L Su, J Zhang, H Meng, T Ouyang, J Li, T Wang, Z Fan, T Fan, B Lin, Y Xie
The prevalence of BRCA1/2 large genomic rearrangements (LGRs) and their underlying mechanisms have not been fully evaluated in Chinese women with breast cancer. In this study, we determined the prevalence of BRCA1/2 LGRs in 834 patients with familial breast cancer and 660 patients with sporadic triple-negative breast cancer who were negative for BRCA1/2 small-range mutations using the multiplex ligation-dependent probe amplification method. We found that twenty index patients (2.4%) in the familial breast cancer group carried a BRCA1 or BRCA2 LGR, and the frequencies of BRCA1 and BRCA2 LGRs were 1...
March 27, 2018: Clinical Genetics
https://www.readbyqxmd.com/read/29578506/crispr-cas9-mediated-targeted-integration-in-vivo-using-a-homology-mediated-end-joining-based-strategy
#6
Xuan Yao, Xing Wang, Junlai Liu, Linyu Shi, Pengyu Huang, Hui Yang
As a promising genome editing platform, the CRISPR/Cas9 system has great potential for efficient genetic manipulation, especially for targeted integration of transgenes. However, due to the low efficiency of homologous recombination (HR) and various indel mutations of non-homologous end joining (NHEJ)-based strategies in non-dividing cells, in vivo genome editing remains a great challenge. Here, we describe a homology-mediated end joining (HMEJ)-based CRISPR/Cas9 system for efficient in vivo precise targeted integration...
March 12, 2018: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/29577652/preliminary-study-of-the-homologous-recombination-repair-pathway-in-mouse-spermatogonial-stem-cells
#7
W Le, L Qi, C Xu, Z Xiang, Z Mao, J Zhang, J Xu, D Wu
The present study was designed to detect DNA repair response through the homologous recombination pathway in mouse spermatogonial stem cells. Mouse spermatogonial stem cells (mSSCs) were obtained from the adult DBA/2 mouse testes by MACS sorting. mSSCs and mice animals were divided into four groups (30 min, 2, 24 h, control) and treated with ionizing irradiation while the control group received pseudo-irradiation. Proteins involved in the homologous recombination pathway (γH2AX, ATM, RAD51, CtIP, and RPA2) were assessed in mSSCs both in vitro and in vivo...
March 25, 2018: Andrology
https://www.readbyqxmd.com/read/29566071/improving-the-efficiency-of-homologous-recombination-by-chemical-and-biological-approaches-in-yarrowia-lipolytica
#8
In-Seung Jang, Byung Jo Yu, Ji Yeon Jang, Jonggeon Jegal, Ju Young Lee
Gene targeting is a challenge in Yarrowia lipolytica (Y. lipolytica) where non-homologous end-joining (NHEJ) is predominant over homologous recombination (HR). To improve the frequency and efficiency of HR in Y. lipolytica, the ku70 gene responsible for a double stand break (DSB) repair in the NHEJ pathway was disrupted, and the cell cycle was synchronized to the S-phase with hydroxyurea, respectively. Consequently, the HR frequency was over 46% with very short homology regions (50 bp): the pex10 gene was accurately deleted at a frequency of 60% and the β-carotene biosynthetic genes were integrated at the correct locus at an average frequency of 53%...
2018: PloS One
https://www.readbyqxmd.com/read/29557773/a-highly-efficient-genetic-system-for-the-identification-of-a-harzianum-b-biosynthetic-gene-cluster-in-trichoderma-hypoxylon
#9
Huan Liu, Gang Wang, Wei Li, Xingzhong Liu, Erwei Li, Wen-Bing Yin
Trichoderma hypoxylon is a fungicolous species which produces rich secondary metabolites. However, no genetic transformation method is available for further studies. Here, we developed a marker-less transformation system based on the complementation of an uridine/uracil biosynthetic gene by protoplast transformation. An uridine/uracil auxotrophic mutant of Δthpyr4 was obtained by using a positive screening protocol with 5'-fluoroorotic acid as a selective reagent. To improve the homologous integration rates, the orthologues of ku70 and lig4 which play critical roles in non-homologous end-joining recombination were disrupted...
March 19, 2018: Microbiology
https://www.readbyqxmd.com/read/29545602/lrh1-enhances-cell-resistance-to-chemotherapy-by-transcriptionally-activating-mdc1-expression-and-attenuating-dna-damage-in-human-breast-cancer
#10
S Wang, Z Zou, X Luo, Y Mi, H Chang, D Xing
Liver receptor homolog-1 (LRH1) has been shown to promote tumor proliferation and development. However, the functions of LRH1 in mediating cancer cells chemoresistance are still not clear. Here, we found LRH1 levels were significantly elevated in primary breast cancer tissues in patients who developed early recurrence. Similarly, adriamycin (ADR)-resistant breast cancer cell lines also exerted high LRH1 expression. Indeed, overexpression of LRH1 attenuated cytotoxicity of chemotherapeutic drugs ADR and cisplatin (DDP) in breast cancer cells in vitro and in nude mice tumor model...
March 16, 2018: Oncogene
https://www.readbyqxmd.com/read/29545335/mutant-idh1-cooperates-with-atrx-loss-to-drive-the-alternative-lengthening-of-telomere-alt-phenotype-in-glioma
#11
Joydeep Mukherjee, Tor-Christian Aase Johannessen, Shigeo Ohba, Tracy T Chow, Lindsey E Jones, Ajay Pandita, Russell O Pieper
A subset of tumors use a recombination-based alternative lengthening of telomere (ALT) pathway to resolve telomeric dysfunction in the absence of TERT. Loss-of-function mutations in the chromatin remodeling factor ATRX are associated with ALT but are insufficient to drive the process. Because many ALT tumors express the mutant isocitrate dehydrogenase IDH1 R132H, including all lower-grade astrocytomas (LGA) and secondary glioblastoma, we examined an hypothesized role for IDH1 R132H in driving the ALT phenotype during gliomagenesis...
March 15, 2018: Cancer Research
https://www.readbyqxmd.com/read/29531807/nf-%C3%AE%C2%BAb-inhibition-by-dimethylaminoparthenolide-radiosensitizes-non-small-cell-lung-carcinoma-by-blocking-dna-double-strand-break-repair
#12
Peter V Deraska, Colin O'Leary, Hunter D Reavis, Shelby Labe, Tru-Khang Dinh, Jean-Bernard Lazaro, Christopher Sweeney, Alan D D'Andrea, David Kozono
Despite optimal chemotherapy, radiotherapy (RT), and/or surgery, non-small-cell lung carcinoma (NSCLC) remains the leading cause of cancer-related death in the US and worldwide. Thoracic RT, a mainstay in the treatment of locally advanced NSCLC, is often restricted in efficacy by a therapeutic index limited by sensitivity of tissues surrounding the malignancy. Therefore, radiosensitizers that can improve the therapeutic index are a vital unmet need. Inhibition of the NF-κB pathway is a proposed mechanism of radiosensitization...
December 2018: Cell Death Discovery
https://www.readbyqxmd.com/read/29527968/linp1-facilitates-dna-damage-repair-through-non-homologous-end-joining-nhej-pathway-and-subsequently-decreases-the-sensitivity-of-cervical-cancer-cells-to-ionizing-radiation
#13
Xuanxuan Wang, Hai Liu, Liming Shi, Xiaoli Yu, Yanjun Gu, Xiaonan Sun
LncRNA in non-homologous end joining (NHEJ) pathway 1 (LINP1) is an lncRNA which promotes therapeutic resistance in triple-negative breast cancer (TNBC). However, the expression and function of LINP1 in cervical cancer is not yet well-understood. In this study, we evaluated the expression levels of LINP1 in tumor tissues and cell lines of cervical cancer. We found that LINP1 associates with NHEJ proteins (Ku80 and DNA-PKcs). LINP1 translocates from cytosol to nucleus in response to irradiation. In addition, LINP1 knockdown significantly increases the levels of cleaved caspase3 and PARP, leading to enhanced cell apoptosis after ionizing radiation (IR)...
March 12, 2018: Cell Cycle
https://www.readbyqxmd.com/read/29522917/dna-damage-response-following-x-irradiation-in-oral-cancer-cell-lines-hsc3-and-hsc4
#14
Sirimanas Jiaranuchart, Atsushi Kaida, Yusuke Onozato, Hiroyuki Harada, Masahiko Miura
OBJECTIVE: The objective of this study was to characterize the DNA damage response in two human oral cancer cell lines following X-irradiation. DESIGN: To visualize radiation-induced cell cycle alterations, two human oral cancer cell lines, HSC3 and HSC4, expressing fluorescent ubiquitination-based cell cycle indicator (Fucci) were established in this study. G2 arrest kinetics following irradiation were obtained from two-color flow cytometric analysis and pedigrees of Fucci fluorescence...
March 6, 2018: Archives of Oral Biology
https://www.readbyqxmd.com/read/29520062/the-h2b-deubiquitinase-usp22-promotes-antibody-class-switch-recombination-by-facilitating-non-homologous-end-joining
#15
Conglei Li, Thergiory Irrazabal, Clare C So, Maribel Berru, Likun Du, Evelyn Lam, Alexanda K Ling, Jennifer L Gommerman, Qiang Pan-Hammarström, Alberto Martin
Class switch recombination (CSR) has a fundamental function during humoral immune response and involves the induction and subsequent repair of DNA breaks in the immunoglobulin (Ig) switch regions. Here we show the role of Usp22, the SAGA complex deubiquitinase that removes ubiquitin from H2B-K120, in the repair of programmed DNA breaks in vivo. Ablation of Usp22 in primary B cells results in defects in γH2AX and impairs the classical non-homologous end joining (c-NHEJ), affecting both V(D)J recombination and CSR...
March 8, 2018: Nature Communications
https://www.readbyqxmd.com/read/29514250/pseudomonas-aeruginosa-mutl-promotes-large-chromosomal-deletions-through-non-homologous-end-joining-to-prevent-bacteriophage-predation
#16
Mengyu Shen, Huidong Zhang, Wei Shen, Zhenyu Zou, Shuguang Lu, Gang Li, Xuesong He, Melissa Agnello, Wenyuan Shi, Fuquan Hu, Shuai Le
Pseudomonas aeruginosa is an opportunistic pathogen with a relatively large genome, and has been shown to routinely lose genomic fragments during environmental selection. However, the underlying molecular mechanisms that promote chromosomal deletion are still poorly understood. In a recent study, we showed that by deleting a large chromosomal fragment containing two closely situated genes, hmgA and galU, P. aeruginosa was able to form 'brown mutants', bacteriophage (phage) resistant mutants with a brown color phenotype...
March 5, 2018: Nucleic Acids Research
https://www.readbyqxmd.com/read/29511339/p73-coordinates-with-%C3%AE-133p53-to-promote-dna-double-strand-break-repair
#17
Hongjian Gong, Yuxi Zhang, Kunpeng Jiang, Shengfan Ye, Shuming Chen, Qinghe Zhang, Jinrong Peng, Jun Chen
Tumour repressor p53 isoform Δ133p53 is a target gene of p53 and an antagonist of p53-mediated apoptotic activity. We recently demonstrated that Δ133p53 promotes DNA double-strand break (DSB) repair by upregulating transcription of the repair genes RAD51, LIG4 and RAD52 in a p53-independent manner. However, Δ133p53 lacks the transactivation domain of full-length p53, and the mechanism by which it exerts transcriptional activity independently of full-length p53 remains unclear. In this report, we describe the accumulation of high levels of both Δ133p53 and p73 (a p53 family member) at 24 h post γ-irradiation (hpi)...
March 6, 2018: Cell Death and Differentiation
https://www.readbyqxmd.com/read/29502733/metformin-prevention-of-genomic-instability-and-cancer-a-review
#18
Masoud Najafi, Mohsen Cheki, Saeed Rezapoor, Ghazale Geraily, Elahe Motevaseli, Carla Carnovale, Emilio Clementi, Alireza Shirazi
The diabetes drug metformin can mitigate the genotoxic effects of cytotoxic agents and has been proposed to prevent or even cure certain cancers. Metformin reduces DNA damage by mechanisms that are only incompletely understood. Metformin scavenges free radicals, including reactive oxygen species and nitric oxide, which are produced by genotoxicants such as ionizing or non-ionizing radiation, heavy metals, and chemotherapeutic agents. The drug may also increase the activities of antioxidant enzymes and inhibit NADPH oxidase, cyclooxygenase-2, and inducible nitric oxide synthase, thereby limiting macrophage recruitment and inflammatory responses...
March 2018: Mutation Research
https://www.readbyqxmd.com/read/29500194/an-efficient-platform-for-generating-somatic-point-mutations-with-germline-transmission-in-the-zebrafish-by-crispr-cas9-mediated-gene-editing
#19
Yibo Zhang, Zhiwei Zhang, Wei Ge
Homology-directed recombination (HDR)-mediated genome editing is a powerful approach for both basic functional study and disease modeling. Although some studies have reported HDR-mediated precise editing in non-rodent models, the efficiency of establishing pure mutant animal lines that carry specific amino acid substitutions remains low. Furthermore, because the efficiency of nonhomologous end joining (NHEJ)-induced insertion and deletion (indel) mutations is normally much higher than that of HDR-induced point mutations, it is often difficult to identify the latter in the background of indel mutations...
March 2, 2018: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/29500182/ku-dna-end-binding-activity-promotes-repair-fidelity-and-influences-end-processing-during-nonhomologous-end-joining-in-saccharomyces-cerevisiae
#20
Charlene H Emerson, Christopher R Lopez, Albert Ribes-Zamora, Erica J Polleys, Christopher L Williams, Lythou Yeo, Jacques E Zaneveld, Rui Chen, Alison A Bertuch
The Ku heterodimer acts centrally in non-homologous end-joining (NHEJ) of DNA double strand breaks (DSB). Saccharomyces cerevisiae Ku, like mammalian Ku, binds and recruits NHEJ factors to DSB ends. Consequently, NHEJ is virtually absent in yeast Ku null ( yku 70Δ or yku80Δ ) strains. Previously, we unexpectedly observed imprecise NHEJ proficiency in a yeast Ku mutant with impaired DNA end-binding (DEB). However, how DEB impairment supported imprecise NHEJ was unknown. Here, we found imprecise NHEJ proficiency to be a feature of a panel of DEB-impaired Ku mutants and that DEB impairment resulted in a deficiency in precise NHEJ...
March 2, 2018: Genetics
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