Abstract
Trinucleotide repeats are a particular class of microsatellites whose large expansions are responsible for at least two dozen human neurological and developmental disorders. Slippage of the two complementary DNA strands during replication, homologous recombination or DNA repair is generally accepted as a mechanism leading to repeat length changes, creating expansions and contractions of the repeat tract. The present review focuses on recent developments on double-strand break repair involving trinucleotide repeat tracts. Experimental evidences in model organisms show that gene conversion and break-induced replication may lead to large repeat tract expansions, while frequent contractions occur either by single-strand annealing between repeat ends or by gene conversion, triggering near-complete contraction of the repeat tract. In the second part of this review, different therapeutic approaches using highly specific single- or double-strand endonucleases targeted to trinucleotide repeat loci are compared. Relative efficacies and specificities of these nucleases will be discussed, as well as their potential strengths and weaknesses for possible future gene therapy of these dramatic disorders.
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Abbreviations
- BIR:
-
Break-induced replication
- CRISPR:
-
Clustered regularly interspaced short palindromic repeats
- SSA:
-
Single-strand annealing
- ZFN:
-
Zinc-finger nucleases
- TALEN:
-
Transcription activator-like effector nuclease
- DSB:
-
Double-strand break
- SDSA:
-
Synthesis-dependent strand annealing
- UAS:
-
Upstream activating sequence
- MRX complex:
-
Mre11-Rad50-Xrs2 complex
- PAM:
-
Protospacer adjacent motif
- iPSC:
-
Induced pluripotent stem cells
- sgRNA (or gRNA):
-
Single-guide RNA
- SpCas9:
-
Streptococcus pyogenes Cas9
- SaCas9:
-
Staphylococcus aureus Cas9
- HNH:
-
Homing endonuclease domain
- HEK293:
-
Human embryonic kidney cell line 293
- K562:
-
Human immortalized myelogenous leukemia cell line
- AAV:
-
Adenovirus-associated vector
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Acknowledgements
The authors wish to thank the continuous support of the Institut Pasteur and of the Centre National de la Recherche Scientifique (CNRS). L. P. is the recipient of a Cifre PhD fellowship from Sanofi. V. M. was the recipient of two post-doctoral fellowships from Fondation Guy Nicolas and from Fondation Hardy.
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Communicated by M. Kupiec.
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Mosbach, V., Poggi, L. & Richard, GF. Trinucleotide repeat instability during double-strand break repair: from mechanisms to gene therapy. Curr Genet 65, 17–28 (2019). https://doi.org/10.1007/s00294-018-0865-1
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DOI: https://doi.org/10.1007/s00294-018-0865-1