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Precision genome editing in the CRISPR era.
Biochem Cell Biol. 2017 Apr; 95(2):187-201.BC

Abstract

With the introduction of precision genome editing using CRISPR-Cas9 technology, we have entered a new era of genetic engineering and gene therapy. With RNA-guided endonucleases, such as Cas9, it is possible to engineer DNA double strand breaks (DSB) at specific genomic loci. DSB repair by the error-prone non-homologous end-joining (NHEJ) pathway can disrupt a target gene by generating insertions and deletions. Alternatively, Cas9-mediated DSBs can be repaired by homology-directed repair (HDR) using an homologous DNA repair template, thus allowing precise gene editing by incorporating genetic changes into the repair template. HDR can introduce gene sequences for protein epitope tags, delete genes, make point mutations, or alter enhancer and promoter activities. In anticipation of adapting this technology for gene therapy in human somatic cells, much focus has been placed on increasing the fidelity of CRISPR-Cas9 and increasing HDR efficiency to improve precision genome editing. In this review, we will discuss applications of CRISPR technology for gene inactivation and genome editing with a focus on approaches to enhancing CRISPR-Cas9-mediated HDR for the generation of cell and animal models, and conclude with a discussion of recent advances and challenges towards the application of this technology for gene therapy in humans.

Authors+Show Affiliations

a Department of Pathology, Dalhousie University, Halifax, NS B3H 4R2, Canada.a Department of Pathology, Dalhousie University, Halifax, NS B3H 4R2, Canada. b Department of Biochemistry & Molecular Biology, Dalhousie University, Halifax, NS B3H 4R2, Canada. c Beatrice Hunter Cancer Research Institute, Halifax, NS B3H 4R2, Canada.

Pub Type(s)

Journal Article
Review

Language

eng

PubMed ID

28177771

Citation

Salsman, Jayme, and Graham Dellaire. "Precision Genome Editing in the CRISPR Era." Biochemistry and Cell Biology = Biochimie Et Biologie Cellulaire, vol. 95, no. 2, 2017, pp. 187-201.
Salsman J, Dellaire G. Precision genome editing in the CRISPR era. Biochem Cell Biol. 2017;95(2):187-201.
Salsman, J., & Dellaire, G. (2017). Precision genome editing in the CRISPR era. Biochemistry and Cell Biology = Biochimie Et Biologie Cellulaire, 95(2), 187-201. https://doi.org/10.1139/bcb-2016-0137
Salsman J, Dellaire G. Precision Genome Editing in the CRISPR Era. Biochem Cell Biol. 2017;95(2):187-201. PubMed PMID: 28177771.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Precision genome editing in the CRISPR era. AU - Salsman,Jayme, AU - Dellaire,Graham, Y1 - 2016/09/29/ PY - 2017/2/9/pubmed PY - 2017/5/26/medline PY - 2017/2/9/entrez KW - CRISPR KW - Cas9 KW - DNA repair KW - gene therapy KW - genome editing KW - réparation d’ADN KW - thérapie génique KW - édition génique SP - 187 EP - 201 JF - Biochemistry and cell biology = Biochimie et biologie cellulaire JO - Biochem Cell Biol VL - 95 IS - 2 N2 - With the introduction of precision genome editing using CRISPR-Cas9 technology, we have entered a new era of genetic engineering and gene therapy. With RNA-guided endonucleases, such as Cas9, it is possible to engineer DNA double strand breaks (DSB) at specific genomic loci. DSB repair by the error-prone non-homologous end-joining (NHEJ) pathway can disrupt a target gene by generating insertions and deletions. Alternatively, Cas9-mediated DSBs can be repaired by homology-directed repair (HDR) using an homologous DNA repair template, thus allowing precise gene editing by incorporating genetic changes into the repair template. HDR can introduce gene sequences for protein epitope tags, delete genes, make point mutations, or alter enhancer and promoter activities. In anticipation of adapting this technology for gene therapy in human somatic cells, much focus has been placed on increasing the fidelity of CRISPR-Cas9 and increasing HDR efficiency to improve precision genome editing. In this review, we will discuss applications of CRISPR technology for gene inactivation and genome editing with a focus on approaches to enhancing CRISPR-Cas9-mediated HDR for the generation of cell and animal models, and conclude with a discussion of recent advances and challenges towards the application of this technology for gene therapy in humans. SN - 1208-6002 UR - https://www.unboundmedicine.com/medline/citation/28177771/Precision_genome_editing_in_the_CRISPR_era_ L2 - https://cdnsciencepub.com/doi/10.1139/bcb-2016-0137?url_ver=Z39.88-2003&rfr_id=ori:rid:crossref.org&rfr_dat=cr_pub=pubmed DB - PRIME DP - Unbound Medicine ER -