scholarly journals Differential efficacies of Cas nucleases on microsatellites involved in human disorders and associated off-target mutations

2019 ◽  
Author(s):  
Lucie Poggi ◽  
Lisa Emmenegger ◽  
Stéphane Descorps-Declère ◽  
Bruno Dumas ◽  
Guy-Franck Richard
Keyword(s):  
Gene Editing ◽  
Low Frequency ◽  
Strand Break ◽  
Double Strand Break ◽  
Yeast Genome ◽  
Guide Rna ◽  
Human Disorders ◽  
Dna End Resection ◽  
End Resection ◽  
Target Effects

AbstractMicrosatellite expansions are the cause of more than 20 neurological or developmental human disorders. Shortening expanded repeats using specific DNA endonucleases may be envisioned as a gene editing approach. Here, a new assay was developed to test several CRISPR-Cas nucleases on microsatellites involved in human diseases, by measuring at the same time double-strand break rates, DNA end resection and homologous recombination efficacy. Broad variations in nuclease performances were detected on all repeat tracts. Streptococcus pyogenes Cas9 was the most efficient of all. All repeat tracts did inhibit double-strand break resection. We demonstrate that secondary structure formation on the guide RNA was a major determinant of nuclease efficacy. Using deep sequencing, off-target mutations were assessed genomewide. Out of 221 CAG/CTG or GAA/TTC trinucleotide repeats of the yeast genome, three were identified as carrying statistically significant low frequency mutations, corresponding to off-target effects.

scholarly journals Regulation of DNA-End Resection by hnRNPU-like Proteins Promotes DNA Double-Strand Break Signaling and Repair

2012 ◽  
Vol 45 (4) ◽  
pp. 505-516 ◽  
Author(s):  
Sophie E. Polo ◽  
Andrew N. Blackford ◽  
J. Ross Chapman ◽  
Linda Baskcomb ◽  
Serge Gravel ◽  
...  
Keyword(s):  
Strand Break ◽  
Double Strand Break ◽  
Dna Double Strand Break ◽  
Dna End Resection ◽  
End Resection

scholarly journals The Effect of Atypical Nucleic Acids Structures in DNA Double Strand Break Repair: A Tale of R-loops and G-Quadruplexes

2021 ◽  
Vol 12 ◽  
Author(s):  
Rosa Camarillo ◽  
Sonia Jimeno ◽  
Pablo Huertas
Keyword(s):  
Nucleic Acids ◽  
Strand Break ◽  
Double Strand Break ◽  
Fine Tuning ◽  
Double Strand Break Repair ◽  
Dna Double Strand Breaks ◽  
Dna Double Strand Break ◽  
Break Repair ◽  
Dna End Resection ◽  
End Resection

The fine tuning of the DNA double strand break repair pathway choice relies on different regulatory layers that respond to environmental and local cues. Among them, the presence of non-canonical nucleic acids structures seems to create challenges for the repair of nearby DNA double strand breaks. In this review, we focus on the recently published effects of G-quadruplexes and R-loops on DNA end resection and homologous recombination. Finally, we hypothesized a connection between those two atypical DNA structures in inhibiting the DNA end resection step of HR.

scholarly journals End resection: a key step in homologous recombination and DNA double-strand break repair

2020 ◽  
Author(s):  
Sijie Liu ◽  
Daochun Kong
Keyword(s):  
Homologous Recombination ◽  
Strand Break ◽  
Double Strand Break ◽  
General Process ◽  
Dna Double Strand Break ◽  
Biochemical Mechanisms ◽  
Strand Invasion ◽  
Dna End Resection ◽  
End Resection ◽  
Made In

AbstractDNA end resection in eukaryotes is a key step in DNA homologous recombination (HR) and HR-mediated DNA double-strand break (DSB) repair, in which DNA2, EXO1 and MRE11 endo- and exonucleases remove several kilobases from the 5′ terminus of the DNA with DSB, while the 3′ terminus remains intact. The end resection-generated 3′ single-stranded DNA (ssDNA) overhang is then coated by RAD51 for subsequent strand invasion. In the last two decades, great progress has been made in understanding the biochemical mechanisms of end resection, including the identification of various enzymes involved in this process. However, some important questions about this process remain to be resolved. In this review, we summarize the general process of end resection and discuss the implications of the most recent findings for understanding of the end resection machinery.

scholarly journals Prolyl Isomerase PIN1 Regulates DNA Double-Strand Break Repair by Counteracting DNA End Resection

2013 ◽  
Vol 50 (3) ◽  
pp. 333-343 ◽  
Author(s):  
Martin Steger ◽  
Olga Murina ◽  
Daniela Hühn ◽  
Lorenza P. Ferretti ◽  
Reto Walser ◽  
...  
Keyword(s):  
Strand Break ◽  
Double Strand Break ◽  
Double Strand Break Repair ◽  
Prolyl Isomerase ◽  
Dna Double Strand Break ◽  
Break Repair ◽  
Dna End Resection ◽  
End Resection

scholarly journals DNA end resection and its role in DNA replication and DSB repair choice in mammalian cells

2020 ◽  
Vol 52 (10) ◽  
pp. 1705-1714 ◽  
Author(s):  
Fei Zhao ◽  
Wootae Kim ◽  
Jake A. Kloeber ◽  
Zhenkun Lou
Keyword(s):  
Dna Repair ◽  
Dna Replication ◽  
Mammalian Cells ◽  
Strand Break ◽  
Double Strand Break ◽  
Double Strand Break Repair ◽  
General Process ◽  
Break Repair ◽  
Dna End Resection ◽  
End Resection

Abstract DNA end resection has a key role in double-strand break repair and DNA replication. Defective DNA end resection can cause malfunctions in DNA repair and replication, leading to greater genomic instability. DNA end resection is initiated by MRN-CtIP generating short, 3′-single-stranded DNA (ssDNA). This newly generated ssDNA is further elongated by multiple nucleases and DNA helicases, such as EXO1, DNA2, and BLM. Effective DNA end resection is essential for error-free homologous recombination DNA repair, the degradation of incorrectly replicated DNA and double-strand break repair choice. Because of its importance in DNA repair, DNA end resection is strictly regulated. Numerous mechanisms have been reported to regulate the initiation, extension, and termination of DNA end resection. Here, we review the general process of DNA end resection and its role in DNA replication and repair pathway choice.

scholarly journals Exo1 plays a major role in DNA end resection in humans and influences double-strand break repair and damage signaling decisions

DNA Repair ◽  
2012 ◽  
Vol 11 (4) ◽  
pp. 441-448 ◽  
Author(s):  
Nozomi Tomimatsu ◽  
Bipasha Mukherjee ◽  
Katherine Deland ◽  
Akihiro Kurimasa ◽  
Emma Bolderson ◽  
...  
Keyword(s):  
Strand Break ◽  
Double Strand Break ◽  
Double Strand Break Repair ◽  
Break Repair ◽  
Dna End Resection ◽  
End Resection
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