Tags

Type your tag names separated by a space and hit enter

Optimization of genome editing through CRISPR-Cas9 engineering.
Bioengineered. 2016 Apr; 7(3):166-74.B

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.

Authors+Show Affiliations

a Metabolic Diseases Branch , National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health , Bethesda , MD , USA.a Metabolic Diseases Branch , National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health , Bethesda , MD , USA.a Metabolic Diseases Branch , National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health , Bethesda , MD , USA.a Metabolic Diseases Branch , National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health , Bethesda , MD , USA.a Metabolic Diseases Branch , National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health , Bethesda , MD , USA.

Pub Type(s)

Journal Article
Review

Language

eng

PubMed ID

27340770

Citation

Zhang, Jian-Hua, et al. "Optimization of Genome Editing Through CRISPR-Cas9 Engineering." Bioengineered, vol. 7, no. 3, 2016, pp. 166-74.
Zhang JH, Adikaram P, Pandey M, et al. Optimization of genome editing through CRISPR-Cas9 engineering. Bioengineered. 2016;7(3):166-74.
Zhang, J. H., Adikaram, P., Pandey, M., Genis, A., & Simonds, W. F. (2016). Optimization of genome editing through CRISPR-Cas9 engineering. Bioengineered, 7(3), 166-74. https://doi.org/10.1080/21655979.2016.1189039
Zhang JH, et al. Optimization of Genome Editing Through CRISPR-Cas9 Engineering. Bioengineered. 2016;7(3):166-74. PubMed PMID: 27340770.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Optimization of genome editing through CRISPR-Cas9 engineering. AU - Zhang,Jian-Hua, AU - Adikaram,Poorni, AU - Pandey,Mritunjay, AU - Genis,Allison, AU - Simonds,William F, PY - 2016/6/25/entrez PY - 2016/6/25/pubmed PY - 2017/1/17/medline KW - CRISPR-Cas9 KW - efficiency KW - genome editing KW - specificity SP - 166 EP - 74 JF - Bioengineered JO - Bioengineered VL - 7 IS - 3 N2 - 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. SN - 2165-5987 UR - https://www.unboundmedicine.com/medline/citation/27340770/Optimization_of_genome_editing_through_CRISPR_Cas9_engineering_ L2 - https://www.tandfonline.com/doi/full/10.1080/21655979.2016.1189039 DB - PRIME DP - Unbound Medicine ER -