Is Non-Homologous End-Joining Really an Inherently Error-Prone Process?

Bétermier, Mireille; Bertrand, Pascale; Lopez, Bernard S.

doi:10.1371/journal.pgen.1004086

Cited by 371 publications

(352 citation statements)

References 98 publications

(132 reference statements)

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“…However, it is important to keep in mind that the CRISPR-Cas9 approach is a nuclease system that induces blunt-ended DSBs. A recent report suggests that the repair of signal joints, which results from the direct ligation of blunt ends, is largely errorfree 28 . The authors also indicate that the accuracy of the repair is dictated by the structure of the DNA ends rather than by the NHEJ machinery.…”

Section: Discussionmentioning

confidence: 99%

Engineering human tumour-associated chromosomal translocations with the RNA-guided CRISPR–Cas9 system

et al. 2014

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Cancer-related human chromosomal translocations are generated through the illegitimate joining of two non-homologous chromosomes affected by double-strand breaks (DSB). Effective methodologies to reproduce precise reciprocal tumour-associated chromosomal translocations are required to gain insight into the initiation of leukaemia and sarcomas. Here we present a strategy for generating cancer-related human chromosomal translocations in vitro based on the ability of the RNA-guided CRISPR-Cas9 system to induce DSBs at defined positions. Using this approach we generate human cell lines and primary cells bearing chromosomal translocations resembling those described in acute myeloid leukaemia and Ewing's sarcoma at high frequencies. FISH and molecular analysis at the mRNA and protein levels of the fusion genes involved in these engineered cells reveal the reliability and accuracy of the CRISPR-Cas9 approach, providing a powerful tool for cancer studies.

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Section: Discussionmentioning

confidence: 99%

Engineering human tumour-associated chromosomal translocations with the RNA-guided CRISPR–Cas9 system

et al. 2014

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“…Classical NHEJ (C-NHEJ), although mainly conservative, can occasionally be associated with point mutations and small deletions, depending on the structure of the DNA ends. Microhomology Mediated End Joining (an alternative NHEJ pathway) leads to the deletion of the sequence between microhomologies (reviewed in 2 ). In addition, recent studies suggest that NHEJ is the primary cause of translocations 3,4 and dysfunctional telomeres fusion.…”

Section: Homologous Recombination and Non Homologous Endmentioning

confidence: 99%

DNA double strand break repair pathway choice: a chromatin based decision?

Clouaire

Legube

2015

Nucleus

100

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“…NHEJ is error-prone as it predominantly ligates broken DNA ends (Betermier et al 2014), whereas the HR pathway is more accurate as it uses an intact copy of the affected region as a template for repair. The a1-a2 repressor expressed in diploid and SIR2 null strains represses specific genes involved in the NHEJ pathway (Åström et al 1999;Valencia et al 2001).…”

mentioning

confidence: 99%

Bypassing the Requirement for an Essential MYST Acetyltransferase

Torres-Machorro

Pillus

2014

Genetics

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Histone acetylation is a key regulatory feature for chromatin that is established by opposing enzymatic activities of lysine acetyltransferases (KATs/HATs) and deacetylases (KDACs/HDACs). Esa1, like its human homolog Tip60, is an essential MYST family enzyme that acetylates histones H4 and H2A and other nonhistone substrates. Here we report that the essential requirement for ESA1 in Saccharomyces cerevisiae can be bypassed upon loss of Sds3, a noncatalytic subunit of the Rpd3L deacetylase complex. By studying the esa1Δ sds3Δ strain, we conclude that the essential function of Esa1 is in promoting the cellular balance of acetylation. We demonstrate this by fine-tuning acetylation through modulation of HDACs and the histone tails themselves. Functional interactions between Esa1 and HDACs of class I, class II, and the Sirtuin family define specific roles of these opposing activities in cellular viability, fitness, and response to stress. The fact that both increased and decreased expression of the ESA1 homolog TIP60 has cancer associations in humans underscores just how important the balance of its activity is likely to be for human well-being.T HE genetic information of DNA is packed into chromatin, which is predominantly composed of the H3, H4, H2A, and H2B core histone proteins that together with DNA form nucleosomes, the basic subunits of the genome (Kornberg and Lorch 1999). Chromatin structure regulates many cellular processes including gene expression, DNA replication, DNA damage repair, and recombination (Felsenfeld and Groudine 2003). Nucleosomes themselves are tightly regulated by mobilization and positioning that are mediated by ATP-dependent chromatin-remodeling machines (Rando and Winston 2012) and by multiple types of histone posttranslational modifications that include acetylation and many other marks (Campos and Reinberg 2009).Nucleosome acetylation is a highly dynamic modification that is promoted by HATs and removed by HDACs. HATs are classified by sequence into different families (Allis et al. 2007). ESA1/KAT5 belongs to the widely conserved MYST HAT family, named for its founding members (MOZ-YBF2/ SAS3-SAS2-TIP60) (Lafon et al. 2007). Esa1 is an essential HAT in yeast (Smith et al. 1998;Clarke et al. 1999) and the catalytic subunit of two distinct multi-protein complexes: NuA4 and Piccolo (Boudreault et al. 2003). Notably, the human homolog of Esa1 is Tip60, which is also essential in vertebrates and has been linked to multiple human diseases (Squatrito et al. 2006;Avvakumov and Côté 2007;Lafon et al. 2007), thus increasing the relevance of gaining a deeper understanding of essential HAT functions.Esa1 primarily acetylates H4 and H2A in vivo (Clarke et al. 1999;Lin et al. 2008) and regulates the expression of active protein-encoding genes (Reid et al. 2000;Lin et al. 2008). It plays a crucial role in cell cycle progression and ribosomal DNA (rDNA) silencing (Clarke et al. 1999(Clarke et al. , 2006 and is recruited to DNA double-strand breaks (DSBs) to promote damage repair by acetylati...

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Is Non-Homologous End-Joining Really an Inherently Error-Prone Process?

Cited by 371 publications

References 98 publications

Engineering human tumour-associated chromosomal translocations with the RNA-guided CRISPR–Cas9 system

Engineering human tumour-associated chromosomal translocations with the RNA-guided CRISPR–Cas9 system

DNA double strand break repair pathway choice: a chromatin based decision?

Bypassing the Requirement for an Essential MYST Acetyltransferase

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