Title

Design and Assembly of CRISPR/Cas9 Lentiviral and rAAV Vectors for Targeted Genome Editing

Document Type

Article

Abstract

Clustered regularly interspaced short palindromic repeat (CRISPR/Cas) system has emerged as an extremely useful tool for biological research and as a potential technology for gene therapy approaches. CRISPR/Cas mediated genome editing can be used to easily and efficiently modify endogenous genes in a large variety of cells and organisms. Furthermore, a modified version of the Cas9 nuclease has been developed that can be used for regulation of endogenous gene expression and labeling of genomic loci, among other applications. This chapter provides an introduction to the basis of the technology and a detail protocol for the most classic application: gene inactivation by CRISPR/Cas9 nuclease system from Streptococcus pyogenes. This workflow can be easily adapted for other CRISPR systems and applications.

Keywords

AAV, CRISPR/Cas9, Cas9 nuclease, Gene editing, Gene inactivation, LV, guideRNA

Medical Subject Headings

Animals; Bacterial Proteins (metabolism); CRISPR-Associated Protein 9 (metabolism); CRISPR-Cas Systems; Dependovirus (genetics); Gene Editing (methods); Gene Expression; Genetic Vectors; HEK293 Cells; Humans; Lentivirus (genetics); RNA, Guide (genetics); Rats; Streptococcus pyogenes (enzymology)

Publication Date

1-1-2019

Publication Title

Methods in molecular biology (Clifton, N.J.)

E-ISSN

1940-6029

Volume

1937

First Page

29

Last Page

45

PubMed ID

30706388

Digital Object Identifier (DOI)

10.1007/978-1-4939-9065-8_2

This document is currently not available here.

Share

COinS