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New Developments in CRISPR Technology: Improvements in Specificity and Efficiency

[ Vol. 18 , Issue. 13 ]

Author(s):

Fatemeh Safari, Safar Farajnia*, Younes Ghasemi and Nosratollah Zarghami   Pages 1038 - 1054 ( 17 )

Abstract:


Background: RNA-guided endonuclease as a versatile genome editing technology opened new windows in various fields of biology. The simplicity of this revolutionary technique provides a promising future for its application in a broad range of approaches from functional annotation of genes to diseases, to genetic manipulation and gene therapy. Besides the site-specific activity of Cas9 endonuclease, the unintended cleavage known as off-target effect is still a major challenge for this genome editing technique.

Methods: Various strategies have been developed to resolve this bottleneck including development of new softwares for designing optimized guide RNA (gRNA), engineering Cas9 enzyme, improvement in off-target detection assays, etc.

Results: This review dedicated to discuss on methods that have been used for optimizing Cas9, specificity with the aim of improving this technology for therapeutic applications.

Conclusion: In addition, the applications and novel breakthroughs in the field of CRISPR technology will be described.

Keywords:

CRISPR-Cas systems, guide RNA (gRNA), gene editing, mutation, off-target effect, single-strand specific DNA and RNA endonucleases.

Affiliation:

Department of Medical Biotechnology, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz, Biotechnology Research Center, Tabriz University of Medical Sciences, Tabriz, Department of Pharmaceutical Biotechnology, School of Pharmacy, and Pharmaceutical Sciences Research Centre, Shiraz University of Medical Sciences, Shiraz, Department of Medical Biotechnology, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz

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