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Optimization of genome editing through CRISPR-Cas9 engineering.
- Source :
-
Bioengineered [Bioengineered] 2016 Apr; Vol. 7 (3), pp. 166-74. - Publication Year :
- 2016
-
Abstract
- CRISPR (Clustered Regularly-Interspaced Short Palindromic Repeats)-Cas9 (CRISPR associated protein 9) has rapidly become the most promising genome editing tool with great potential to revolutionize medicine. Through guidance of a 20 nucleotide RNA (gRNA), CRISPR-Cas9 finds and cuts target protospacer DNA precisely 3 base pairs upstream of a PAM (Protospacer Adjacent Motif). The broken DNA ends are repaired by either NHEJ (Non-Homologous End Joining) resulting in small indels, or by HDR (Homology Directed Repair) for precise gene or nucleotide replacement. Theoretically, CRISPR-Cas9 could be used to modify any genomic sequences, thereby providing a simple, easy, and cost effective means of genome wide gene editing. However, the off-target activity of CRISPR-Cas9 that cuts DNA sites with imperfect matches with gRNA have been of significant concern because clinical applications require 100% accuracy. Additionally, CRISPR-Cas9 has unpredictable efficiency among different DNA target sites and the PAM requirements greatly restrict its genome editing frequency. A large number of efforts have been made to address these impeding issues, but much more is needed to fully realize the medical potential of CRISPR-Cas9. In this article, we summarize the existing problems and current advances of the CRISPR-Cas9 technology and provide perspectives for the ultimate perfection of Cas9-mediated genome editing.
- Subjects :
- Animals
Bacterial Proteins metabolism
CRISPR-Associated Protein 9
DNA chemistry
DNA genetics
DNA metabolism
DNA End-Joining Repair
Endonucleases metabolism
Humans
Models, Molecular
Nucleotide Motifs
RNA, Guide, CRISPR-Cas Systems metabolism
Recombinational DNA Repair
Bacterial Proteins genetics
CRISPR-Cas Systems
Clustered Regularly Interspaced Short Palindromic Repeats
Endonucleases genetics
Gene Editing methods
Genome
RNA, Guide, CRISPR-Cas Systems genetics
Subjects
Details
- Language :
- English
- ISSN :
- 2165-5987
- Volume :
- 7
- Issue :
- 3
- Database :
- MEDLINE
- Journal :
- Bioengineered
- Publication Type :
- Academic Journal
- Accession number :
- 27340770
- Full Text :
- https://doi.org/10.1080/21655979.2016.1189039