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CRISPR-Directed Therapeutic Correction at the Locus Is Challenged by Frequent Incidence of Chromosomal Deletions

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Publisher Cell Press
Date 2020 May 19
PMID 32420407
Citations 5
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Abstract

Resurrection of non-processed pseudogenes may increase the efficacy of therapeutic gene editing, upon simultaneous targeting of a mutated gene and its highly homologous pseudogenes. To investigate the potency of this approach for clinical gene therapy of human diseases, we corrected a pseudogene-associated disorder, the immunodeficiency p47 -deficient chronic granulomatous disease (p47 CGD), using clustered regularly interspaced short palindromic repeats-associated nuclease Cas9 (CRISPR-Cas9) to target mutated neutrophil cytosolic factor 1 (). Being separated by less than two million base pairs, and two pseudogenes are closely co-localized on chromosome 7. In healthy people, a two-nucleotide GT deletion (ΔGT) is present in the and pseudogenes only. In the majority of patients with p47 CGD, the gene is inactivated due to a ΔGT transfer from one of the two non-processed pseudogenes. Here we demonstrate that concurrent targeting and correction of mutated and its pseudogenes results in therapeutic CGD phenotype correction, but also causes potentially harmful chromosomal deletions between the targeted loci in a p47 -deficient CGD cell line model. Therefore, development of genome-editing-based treatment of pseudogene-related disorders mandates thorough safety examination, as well as technological advances, limiting concurrent induction of multiple double-strand breaks on a single chromosome.

Citing Articles

Gene editing of NCF1 loci is associated with homologous recombination and chromosomal rearrangements.

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Targeted knock-in of cDNA into the locus leads to myeloid phenotypic correction of p47 -deficient chronic granulomatous disease.

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[Gene editing for the treatment of primary immunodeficiency disease].

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Therapy Development by Genome Editing of Hematopoietic Stem Cells.

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