Author(s): PRADO, G. S.; PINHEIRO, T. T.; FARIA, J. C. de; VIANELLO, R. P.

Summary: We will focus here on the use of the CRISPR/Cas system to induce mutations via NHEJ. The NHEJ mechanism acts in most of the cell cycle and is not based on homology to repair DNA, causing mutations such as insertions or deletions (indels) or even substitutions. In this way, the expression of the genes in the place where the repair occurred by NHEJ can, in general, be interrupted, and the encoded proteins may not be functional. To clarify the process of genome editing by CRISPR, this chapter aims to address the details of the NHEJ mechanism, as well as the use of ribonucleoprotein systems (RNPs) to promote editing, in addition to providing a practical procedure for the design of experiments.

Publication year: 2021

Types of publication: Book sections

Unit: Embrapa Rice & Beans

Keywords: Biotechnology, Biotecnologia, CRISPR, CRISPRevolution, DNA, Engenharia Genética, Genetic engineering, Genoma, Non-homologous, Nucleases, Plant genetics, RNA, RNA editing, Ribonucleoproteins