scholarly journals New Insight on Entangled DNA Repair Pathways: Stable Silenced Human Cells for Unraveling the DDR Jigsaw

10.5772/23681 ◽  
2011 ◽  
Author(s):  
Biard Denis
Keyword(s):  
Dna Repair ◽  
Human Cells ◽  
Repair Pathways

scholarly journals CRISPR-Cas9 genome editing in human cells works via the Fanconi Anemia pathway

2017 ◽  
Author(s):  
Chris D Richardson ◽  
Katelynn R Kazane ◽  
Sharon J Feng ◽  
Nicholas L Bray ◽  
Axel J Schäfer ◽  
...  
Keyword(s):  
Dna Repair ◽  
Genome Editing ◽  
Fanconi Anemia ◽  
High Efficiency ◽  
Genetic Diseases ◽  
Dermal Fibroblasts ◽  
Human Cells ◽  
Individual Gene ◽  
Break Site ◽  
Repair Pathways

AbstractCRISPR-Cas9 genome editing creates targeted double strand breaks (DSBs) in eukaryotic cells that are processed by cellular DNA repair pathways. Co-administration of single stranded oligonucleotide donor DNA (ssODN) during editing can result in high-efficiency (>20%) incorporation of ssODN sequences into the break site. This process is commonly referred to as homology directed repair (HDR) and here referred to as single stranded template repair (SSTR) to distinguish it from repair using a double stranded DNA donor (dsDonor). The high efficacy of SSTR makes it a promising avenue for the treatment of genetic diseases1,2, but the genetic basis of SSTR editing is still unclear, leaving its use a mostly empiric process. To determine the pathways underlying SSTR in human cells, we developed a coupled knockdown-editing screening system capable of interrogating multiple editing outcomes in the context of thousands of individual gene knockdowns. Unexpectedly, we found that SSTR requires multiple components of the Fanconi Anemia (FA) repair pathway, but does not require Rad51-mediated homologous recombination, distinguishing SSTR from repair using dsDonors. Knockdown of FA genes impacts SSTR without altering break repair by non-homologous end joining (NHEJ) in multiple human cell lines and in neonatal dermal fibroblasts. Our results establish an unanticipated and central role for the FA pathway in templated repair from single stranded DNA by human cells. Therapeutic genome editing has been proposed to treat genetic disorders caused by deficiencies in DNA repair, including Fanconi Anemia. Our data imply that patient genotype and/or transcriptome profoundly impact the effectiveness of gene editing treatments and that adjuvant treatments to bias cells towards FA repair pathways could have considerable therapeutic value.

scholarly journals BRCA1 and S phase DNA repair pathways restrict LINE-1 retrotransposition in human cells

2020 ◽  
Vol 27 (2) ◽  
pp. 179-191 ◽  
Author(s):  
Paolo Mita ◽  
Xiaoji Sun ◽  
David Fenyö ◽  
David J. Kahler ◽  
Donghui Li ◽  
...  
Keyword(s):  
Dna Repair ◽  
S Phase ◽  
Human Cells ◽  
Repair Pathways

scholarly journals Preserving genome integrity in human cells via DNA double-strand break repair

2020 ◽  
Vol 31 (9) ◽  
pp. 859-865 ◽  
Author(s):  
Ryan B. Jensen ◽  
Eli Rothenberg
Keyword(s):  
Dna Repair ◽  
Strand Break ◽  
Double Strand Break ◽  
Human Cells ◽  
Cell Cycle Checkpoints ◽  
Human Diseases ◽  
Genome Integrity ◽  
Dsb Repair ◽  
Dna Double Strand Break ◽  
Repair Pathways

The efficient maintenance of genome integrity in the face of cellular stress is vital to protect against human diseases such as cancer. DNA replication, chromatin dynamics, cellular signaling, nuclear architecture, cell cycle checkpoints, and other cellular activities contribute to the delicate spatiotemporal control that cells utilize to regulate and maintain genome stability. This perspective will highlight DNA double-strand break (DSB) repair pathways in human cells, how DNA repair failures can lead to human disease, and how PARP inhibitors have emerged as a novel clinical therapy to treat homologous recombination-deficient tumors. We briefly discuss how failures in DNA repair produce a permissive genetic environment in which preneoplastic cells evolve to reach their full tumorigenic potential. Finally, we conclude that an in-depth understanding of DNA DSB repair pathways in human cells will lead to novel therapeutic strategies to treat cancer and potentially other human diseases.

scholarly journals BRCA1 Mediated Homologous Recombination and S Phase DNA Repair Pathways Restrict LINE-1 Retrotransposition in Human Cells

2019 ◽  
Author(s):  
Xiaoji Sun ◽  
Paolo Mita ◽  
David J. Kahler ◽  
Donghui Li ◽  
Aleksandra Wudzinska ◽  
...  
Keyword(s):  
Dna Repair ◽  
Homologous Recombination ◽  
Potential Role ◽  
Genome Instability ◽  
Protein Translation ◽  
Human Cells ◽  
Host Factors ◽  
Factors Affecting ◽  
A Genome ◽  
Repair Pathways

AbstractLong interspersed element-1 (LINE-1 or L1) is the only autonomous retrotransposon active in human cells. L1s DNA makes about 17% of the human genome and retrotransposition of a few active L1 copies has been detected in various tumors, underscoring the potential role of L1 in mediating or increasing genome instability during tumorigenic development. Different host factors have been shown to influence L1 mobility through several mechanisms. However, systematic analyses of host factors affecting L1 retrotransposition are limited. Here, we developed a high-throughput microscopy-based retrotransposition assay and coupled it to a genome-wide siRNA knockdown screen to study the cellular regulators of L1 retrotransposition in human cells. We showed that L1 insertion frequency was stimulated by knockdown of Double-Stranded Break (DSB) repair factors that are active in the S/G2 phase of the cell cycle including Homologous Recombination (HR), Fanconi Anemia (FA) and, to a less extent, microhomology-mediated end-joining (MMEJ) factors. In particular, we show that BRCA1, an E3 ubiquitin ligase with a key role in several DNA repair pathways, plays multiple roles in regulating L1; BRCA1 knockdown directly affects L1 retrotransposition frequency and structure and also plays a role in controlling L1 ORF2 protein translation through L1 mRNA binding. These results suggest the existence of a “battle” between HR factors and L1 retrotransposons, revealing a potential role for L1 in development of tumors characterized by BRCA1 and HR repair deficiencies.

scholarly journals Targeting DNA Repair and Chromatin Crosstalk in Cancer Therapy

Cancers ◽  
2021 ◽  
Vol 13 (3) ◽  
pp. 381
Author(s):  
Danielle P. Johnson ◽  
Mahesh B. Chandrasekharan ◽  
Marie Dutreix ◽  
Srividya Bhaskara
Keyword(s):  
Dna Repair ◽  
Cancer Therapy ◽  
Therapeutic Intervention ◽  
Synthetic Lethality ◽  
Checkpoint Proteins ◽  
Cellular Processes ◽  
Repair Pathways ◽  
Made In

Aberrant DNA repair pathways that underlie developmental diseases and cancers are potential targets for therapeutic intervention. Targeting DNA repair signal effectors, modulators and checkpoint proteins, and utilizing the synthetic lethality phenomena has led to seminal discoveries. Efforts to efficiently translate the basic findings to the clinic are currently underway. Chromatin modulation is an integral part of DNA repair cascades and an emerging field of investigation. Here, we discuss some of the key advancements made in DNA repair-based therapeutics and what is known regarding crosstalk between chromatin and repair pathways during various cellular processes, with an emphasis on cancer.

17α-ethynilestradiol and tributyltin mixtures modulates the expression of NER and p53 DNA repair pathways in male zebrafish gonads and disrupt offspring embryonic development

2018 ◽  
Vol 95 ◽  
pp. 1008-1018 ◽  
Author(s):  
Joana Soares ◽  
Teresa Neuparth ◽  
Angeliki Lyssimachou ◽  
Daniela Lima ◽  
Ana André ◽  
...  
Keyword(s):  
Dna Repair ◽  
Embryonic Development ◽  
Repair Pathways

Investigation on modulation of DNA repair pathways in Chinese MJD patients

2018 ◽  
Vol 71 ◽  
pp. 267.e5-267.e6 ◽  
Author(s):  
Chunrong Wang ◽  
Zhao Chen ◽  
Huirong Peng ◽  
Yun Peng ◽  
Xin Zhou ◽  
...  
Keyword(s):  
Dna Repair ◽  
Repair Pathways
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