Non-homologous end joining pathway of the human pathogen Cryptococcus neoformans influences homologous integration efficiency but not virulence

Mycoscience ◽  
2010 ◽  
Vol 51 (4) ◽  
pp. 272-280 ◽  
Author(s):  
Hao-Man Li ◽  
Eric V. Virtudazo ◽  
Akio Toh-e ◽  
Masashi Yamaguchi ◽  
Susumu Kawamoto ◽  
...  
Keyword(s):  
Cryptococcus Neoformans ◽  
Human Pathogen ◽  
End Joining ◽  
Homologous Integration ◽  
Non Homologous End Joining ◽  
Integration Efficiency

Increased homologous integration frequency in Yarrowia lipolytica strains defective in non-homologous end-joining

2013 ◽  
Vol 59 (1-2) ◽  
pp. 63-72 ◽  
Author(s):  
Anne Kretzschmar ◽  
Christina Otto ◽  
Martina Holz ◽  
Severine Werner ◽  
Linda Hübner ◽  
...  
Keyword(s):  
Yarrowia Lipolytica ◽  
End Joining ◽  
Homologous Integration ◽  
Non Homologous End Joining

scholarly journals Curing hemophilia A by NHEJ-mediated ectopic F8 insertion in the mouse

2019 ◽  
Vol 20 (1) ◽  
Author(s):  
Jian-Ping Zhang ◽  
Xin-Xin Cheng ◽  
Mei Zhao ◽  
Guo-Hua Li ◽  
Jing Xu ◽  
...  
Keyword(s):  
Mouse Liver ◽  
Hemophilia A ◽  
Fold Increase ◽  
Therapeutic Effects ◽  
End Joining ◽  
Promising Strategy ◽  
Non Homologous End Joining ◽  
Integration Efficiency

Abstract Background Hemophilia A, a bleeding disorder resulting from F8 mutations, can only be cured by gene therapy. A promising strategy is CRISPR-Cas9-mediated precise insertion of F8 in hepatocytes at highly expressed gene loci, such as albumin (Alb). Unfortunately, the precise in vivo integration efficiency of a long insert is very low (~ 0.1%). Results We report that the use of a double-cut donor leads to a 10- to 20-fold increase in liver editing efficiency, thereby completely reconstituting serum F8 activity in a mouse model of hemophilia A after hydrodynamic injection of Cas9-sgAlb and B domain-deleted (BDD) F8 donor plasmids. We find that the integration of a double-cut donor at the Alb locus in mouse liver is mainly through non-homologous end joining (NHEJ)-mediated knock-in. We then target BDDF8 to multiple sites on introns 11 and 13 and find that NHEJ-mediated insertion of BDDF8 restores hemostasis. Finally, using 3 AAV8 vectors to deliver genome editing components, including Cas9, sgRNA, and BDDF8 donor, we observe the same therapeutic effects. A follow-up of 100 mice over 1 year shows no adverse effects. Conclusions These findings lay the foundation for curing hemophilia A by NHEJ knock-in of BDDF8 at Alb introns after AAV-mediated delivery of editing components.

scholarly journals Functional Roles of Homologous Recombination and Non-Homologous End Joining in DNA Damage Response and Microevolution in Cryptococcus neoformans

2021 ◽  
Vol 7 (7) ◽  
pp. 566
Author(s):  
Kwang-Woo Jung ◽  
Jong-Hyun Jung ◽  
Ha-Young Park
Keyword(s):  
Dna Damage ◽  
Homologous Recombination ◽  
Cryptococcus Neoformans ◽  
Dna Damage Response ◽  
Genotoxic Stress ◽  
Dna Lesions ◽  
Dependent Manner ◽  
End Joining ◽  
Damage Response ◽  
Non Homologous End Joining

DNA double-strand breaks (DSBs) are the most deleterious type of DNA lesions because they cause loss of genetic information if not properly repaired. In eukaryotes, homologous recombination (HR) and non-homologous end joining (NHEJ) are required for DSB repair. However, the relationship of HR and NHEJ in DNA damage stress is unknown in the radiation-resistant fungus Cryptococcus neoformans. In this study, we found that the expression levels of HR- and NHEJ-related genes were highly induced in a Rad53–Bdr1 pathway-dependent manner under genotoxic stress. Deletion of RAD51, which is one of the main components in the HR, resulted in growth under diverse types of DNA damage stress, whereas perturbations of KU70 and KU80, which belong to the NHEJ system, did not affect the genotoxic stresses except when bleomycin was used for treatment. Furthermore, deletion of both RAD51 and KU70/80 renders cells susceptible to oxidative stress. Notably, we found that deletion of RAD51 induced a hypermutator phenotype in the fluctuation assay. In contrast to the fluctuation assay, perturbation of KU70 or KU80 induced rapid microevolution similar to that induced by the deletion of RAD51. Collectively, Rad51-mediated HR and Ku70/Ku80-mediated NHEJ regulate the DNA damage response and maintain genome stability.

scholarly journals Withanolide D Enhances Radiosensitivity of Human Cancer Cells by Inhibiting DNA Damage Non-homologous End Joining Repair Pathway

2020 ◽  
Vol 9 ◽  
Author(s):  
Jerome Lacombe ◽  
Titouan Cretignier ◽  
Laetitia Meli ◽  
E. M. Kithsiri Wijeratne ◽  
Jean-Luc Veuthey ◽  
...  
Keyword(s):  
Dna Damage ◽  
Cancer Cells ◽  
Human Cancer ◽  
Repair Pathway ◽  
End Joining ◽  
Human Cancer Cells ◽  
Non Homologous End Joining

scholarly journals Regulatory and Functional Involvement of Long Non-Coding RNAs in DNA Double-Strand Break Repair Mechanisms

Cells ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 1506
Author(s):  
Angelos Papaspyropoulos ◽  
Nefeli Lagopati ◽  
Ioanna Mourkioti ◽  
Andriani Angelopoulou ◽  
Spyridon Kyriazis ◽  
...  
Keyword(s):  
Therapeutic Potential ◽  
Strand Break ◽  
Dna Double Strand Breaks ◽  
End Joining ◽  
Dsb Repair ◽  
Repair Mechanisms ◽  
Non Coding Rnas ◽  
Repair Pathways ◽  
Non Homologous End Joining ◽  
Living Organisms

Protection of genome integrity is vital for all living organisms, particularly when DNA double-strand breaks (DSBs) occur. Eukaryotes have developed two main pathways, namely Non-Homologous End Joining (NHEJ) and Homologous Recombination (HR), to repair DSBs. While most of the current research is focused on the role of key protein players in the functional regulation of DSB repair pathways, accumulating evidence has uncovered a novel class of regulating factors termed non-coding RNAs. Non-coding RNAs have been found to hold a pivotal role in the activation of DSB repair mechanisms, thereby safeguarding genomic stability. In particular, long non-coding RNAs (lncRNAs) have begun to emerge as new players with vast therapeutic potential. This review summarizes important advances in the field of lncRNAs, including characterization of recently identified lncRNAs, and their implication in DSB repair pathways in the context of tumorigenesis.

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