Precision Genome Editing with CRISPR-Cas9

Shahroz Rahman; Abdul Rehman Ikram; Farrukh Azeem; Muhammad Tahir Ul Qamar; Tayyaba Shaheen; Mehboob-Ur-Rahman

doi:10.1007/978-1-0716-3782-1_21

Precision Genome Editing with CRISPR-Cas9

Methods Mol Biol. 2024:2788:355-372. doi: 10.1007/978-1-0716-3782-1_21.

Authors

Shahroz Rahman¹, Abdul Rehman Ikram¹, Farrukh Azeem¹, Muhammad Tahir Ul Qamar¹, Tayyaba Shaheen², Mehboob-Ur-Rahman³

Affiliations

¹ Department of Bioinformatics and Biotechnology, Government College University, Faisalabad, Pakistan.
² Department of Bioinformatics and Biotechnology, Government College University, Faisalabad, Pakistan. tayyabashaheen@edu.gcuf.pk.
³ Plant Genomics and Molecular Breeding Laboratory, National Institute for Biotechnology and Genetic Engineering College, Pakistan Institute of Engineering and Applied Sciences (NIBGE-C, PIEAS), Faisalabad, Punjab, Pakistan. mehboob@nibge.org.

PMID: 38656525
DOI: 10.1007/978-1-0716-3782-1_21

Abstract

The CRISPR/Cas9 system is a revolutionary technology for genome editing that allows for precise and efficient modifications of DNA sequences. The system is composed of two main components, the Cas9 enzyme and a guide RNA (gRNA). The gRNA is designed to specifically target a desired DNA sequence, while the Cas9 enzyme acts as molecular scissors to cut the DNA at that specific location. The cell then repairs the digested DNA, either through nonhomologous end joining (NHEJ) or homology-directed repair (HDR), resulting in either indels or precise modifications of DNA sequences with broad implications in biotechnology, agriculture, and medicine. This chapter provides a practical approach for utilizing CRISPR/Cas9 in precise genome editing, including identifying the target gene sequence, designing gRNA and protein (Cas9), and delivering the CRISPR components to target cells.

Keywords: CRISPR; CRISPR-Cas9; Gene editing; RNP; gRNA.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

CRISPR-Associated Protein 9 / genetics
CRISPR-Associated Protein 9 / metabolism
CRISPR-Cas Systems*
DNA End-Joining Repair
Gene Editing* / methods
Humans
RNA, Guide, CRISPR-Cas Systems* / genetics

Substances

RNA, Guide, CRISPR-Cas Systems
CRISPR-Associated Protein 9