Work

In Vitro Pre-validation of Gene Editing by CRISPR/Cas9 Ribonucleoprotein.

Year: 2019

Type: article

Abstract: BACKGROUND: applications in which the crRNAs in the tracrRNA-crRNA structure guides the Mg2+-dependent RNAdirected DNA endonuclease to introduce double-strand breaks at specific sites in DNA. METHODS: Here, Cas9-crRNA-tracrRNA RNP system was used to test the designed crRNAs for in vitro DNA cleavage by Cas9 protein in RAG1, RAG2 and IL2RG genes. RESULTS: The results of cleavage reveal theCas9-crRNA-tracrRNA RNP system is a rapid and efficient way to pre-validate the efficiency of CRISPR cleavage with crRNAs designed for RAG1, RAG2 and IL2RG genes. CONCLUSION: cleavage of DNA by CRISPR/Cas9 ribonucleoprotein complex can be used to pre-validate the functionality and relative efficiency of CRISPR system for targeting genes.

Source: Europe PMC (PubMed Central)

Authors: Maryam Mehravar, Abolfazl Shirazi, Mohammad Mehdi Mehrazar, Mahboobeh Nazari

Institutions: Academic Center for Education, Culture and Research, Shahrekord University

Language: English

Topic: CRISPR and Genetic Engineering

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: green

OpenAlex ID: W2948411316

FWCI: 1.264

Cites: 22

Cited by: 33

Related to: 10