Molecular basis of xeroderma pigmentosum group C DNA recognition by engineered meganucleases

  title={Molecular basis of xeroderma pigmentosum group C DNA recognition by engineered meganucleases},
  author={Pilar Redondo and Jes{\'u}s Prieto and In{\'e}s G. Mu{\~n}oz and Andreu Alib{\'e}s and François Stricher and Luis Serrano and J P Cabaniols and Fayza Daboussi and Sylvain Arnould and Christophe Perez and Philippe Duchateau and Fr{\'e}d{\'e}ric P{\^a}ques and Francisco J. Blanco and Guillermo Montoya},
Xeroderma pigmentosum is a monogenic disease characterized by hypersensitivity to ultraviolet light. The cells of xeroderma pigmentosum patients are defective in nucleotide excision repair, limiting their capacity to eliminate ultraviolet-induced DNA damage, and resulting in a strong predisposition to develop skin cancers. The use of rare cutting DNA endonucleases—such as homing endonucleases, also known as meganucleases—constitutes one possible strategy for repairing DNA lesions. Homing… 

Molecular basis of engineered meganuclease targeting of the endogenous human RAG1 locus

This is the first time that an engineered meganuclease variant targets the human RAG1 locus by stimulating homologous recombination in human cell lines up to 265 bp away from the cleavage site.

Genetic therapy of Xeroderma Pigmentosum: analysis of strategies and translation

Taking into account the different facets of the disease in its different forms, the potential therapeutic approaches that could help improving life conditions of those rare patients who still lack any efficient cure are discussed.

In vivo genome editing as a potential treatment strategy for inherited retinal dystrophies

Meganucleases and Their Biomedical Applications

This review provides an overview of recent advances in homing endonucleases and zinc-finger nucleases, which are double-stranded DNAses that target large recognition sites and can recognise unique sites of cleavage within a genome, permitting their use for therapeutic purposes in human diseases.

Progress in Retinal and Eye Research

The authors focus on the different DNA repair mechanisms, the current state of the art tools for genome editing and the particularities of the retina and photoreceptors with regard to in vivo therapeutic approaches.

Crystal structure of I-DmoI in complex with its target DNA provides new insights into meganuclease engineering

The crystal structures at 2.0 and 2.1 Å resolution of the I-DmoI meganuclease in complex with its substrate DNA before and after cleavage are reported, providing snapshots of the catalytic process and indicating key residues responsible for nonpalindromic target DNA recognition.

Efficient targeting of a SCID gene by an engineered single-chain homing endonuclease

The first demonstration that an engineered meganuclease can induce targeted recombination at an endogenous locus in up to 6% of transfected human cells is provided, ranking this new generation of endonucleases among the best molecular scissors available for genome surgery strategies.

Crystal Structure of the Homing Endonuclease I-CvuI Provides a New Template for Genome Modification*

The crystal structure of I-CvuI homing endonuclease expands the current repertoire for engineering custom specificities, both by itself as a new scaffold alone and in hybrid constructs with other related homingendonucleases or other DNA-binding protein templates.

Meganucleases can restore the reading frame of a mutated dystrophin

It is a proof of principle that MGNs that are adequately engineered to target appropriate sequences in the human dystrophin gene should be able to restore the normal reading frame of that gene in DMD patients with an out-of-frame deletion.



Clues to epidermal cancer proneness revealed by reconstruction of DNA repair-deficient xeroderma pigmentosum skin in vitro

Compared with normal skin, the DNA repair deficiency of in vitro reconstructed XP skin was documented by long-lasting persistence of UVB-induced DNA damage in all epidermal layers, including the basal layer from which carcinoma develops.

Cancer in xeroderma pigmentosum and related disorders of DNA repair

Nucleotide-excision repair diseases exhibit cancer, complex developmental disorders and neurodegeneration, and complex clinical phenotypes might result from unanticipated effects on other genes and proteins.

Highly efficient endogenous human gene correction using designed zinc-finger nucleases

It is shown that zinc-finger nucleases designed against an X-linked severe combined immune deficiency mutation in the IL2Rγ gene yielded more than 18% gene-modified human cells without selection, raising the possibility of strategies based on zinc- finger nucleases for the treatment of disease.

The C-terminal loop of the homing endonuclease I-CreI is essential for site recognition, DNA binding and cleavage

It is demonstrated that the C-terminal loop of I-CreI endonuclease plays a fundamental role in its catalytic mechanism and suggested this novel site as a region to take into account for engineering new endonucleases with tailored specificity.

A combinatorial approach to create artificial homing endonucleases cleaving chosen sequences

The first artificial HEs whose specificity has been entirely redesigned to cleave a naturally occurring sequence are described, demonstrating that the plasticity of LAGLIDADG endonucleases allows extensive engineering, and provide a general method to create novel endon nucleases with tailored specificities.

An xrcc4 defect or Wortmannin stimulates homologous recombination specifically induced by double-strand breaks in mammalian cells.

A model according to which NHEJ and HR do not simply compete for DSB repair but can act sequentially is suggested: a defect in a late N HEJ step is not a dead end and can make DSB available for subsequent Rad51 recombination complex assembly.

Directed evolution of the site specificity of Cre recombinase

  • S. SantoroP. Schultz
  • Biology
    Proceedings of the National Academy of Sciences of the United States of America
  • 2002
A directed evolution strategy is developed that can be used to identify recombinases that recognize variant loxP sites and selectively recombines a novel recombination site and operates at a rate identical to that of wild-type Cre.