New Developments in CRISPR Technology: Improvements in Specificity and Efficiency

Author(s): Fatemeh Safari, Safar Farajnia*, Younes Ghasemi, Nosratollah Zarghami.

Journal Name: Current Pharmaceutical Biotechnology

Volume 18 , Issue 13 , 2017

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Graphical 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.

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Article Details

Year: 2017
Page: [1038 - 1054]
Pages: 17
DOI: 10.2174/1389201019666180209120533
Price: $65

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