Computational Approaches for Designing Highly Specific and Efficient sgRNAs

Jaspreet Kaur Dhanjal; Dhvani Vora; Navaneethan Radhakrishnan; Durai Sundar

doi:10.1007/978-1-0716-1585-0_8

Computational Approaches for Designing Highly Specific and Efficient sgRNAs

Methods Mol Biol. 2022:2349:147-166. doi: 10.1007/978-1-0716-1585-0_8.

Authors

Jaspreet Kaur Dhanjal¹, Dhvani Vora¹, Navaneethan Radhakrishnan¹, Durai Sundar²

Affiliations

¹ Department of Biochemical Engineering and Biotechnology, DBT-AIST International Laboratory for Advanced Biomedicine (DAILAB), Indian Institute of Technology Delhi, Hauz Khas, New Delhi, India.
² Department of Biochemical Engineering and Biotechnology, DBT-AIST International Laboratory for Advanced Biomedicine (DAILAB), Indian Institute of Technology Delhi, Hauz Khas, New Delhi, India. sundar@dbeb.iitd.ac.in.

PMID: 34718995
DOI: 10.1007/978-1-0716-1585-0_8

Abstract

The easily programmable CRISPR/Cas9 system has found applications in biomedical research as well as microbial and crop applications, due to its ability to create site-specific edits. This powerful and flexible system has also been modified to enable inducible gene regulation, epigenome modifications and high-throughput screens. Designing efficient and specific guides for the nuclease is a key step and also a major challenge in effective application. This chapter describes rules for sgRNA design and important features to consider while touching upon bioinformatics advances in predicting efficient guides. Computational tools that suggest improved guides, depending on application, or predict off-targets have also been mentioned and compared.

Keywords: CRISPR/Cas9; Knockin; Knockout; Off-target; Target; sgRNA.

MeSH terms

CRISPR-Cas Systems / genetics
Computational Biology*
Endonucleases
RNA, Guide, CRISPR-Cas Systems

Substances

RNA, Guide, CRISPR-Cas Systems
Endonucleases