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Genome Editing

Overview

A well-established technique for modifying specific sequences in the genome is gene targeting by homologous recombination, but this method can be laborious and only works in certain organisms. Recent advances have led to the development of “genome editing”, which works by inducing double-strand breaks in DNA using engineered nuclease enzymes guided to target genomic sites by either proteins or RNAs that recognize specific sequences. When a cell attempts to repair this damage, mutations can be introduced into the targeted DNA region.

In this video, JoVE explains the principles behind genome editing, emphasizing how this technique relates to DNA repair mechanisms. Then, three major genome editing methods—zinc finger nucleases, TALENs, and the CRISPR-Cas9 system—are reviewed, followed by a protocol for using CRISPR to create targeted genetic changes in mammalian cells. Finally, we discuss some current research that applies genome editing to alter the genetic material in model organisms or cultured cells.

Procedure

Genome editing comprises techniques with which researchers can “edit” or change a specific DNA sequence. These methods rely on the creation of small “cuts” in DNA, which cells attempt to repair, often incompletely. In this manner, scientists can induce mutations in targeted sequences with greater efficiency compared to classical gene targeting.

In this video, we will review the principles behind three genome editing techniques, and discuss a generalized protocol for one of them, the CRI

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Tags
Genome EditingDNA SequenceSmall Cuts In DNAMutationsGene TargetingCRISPR Cas9 SystemApplicationsHomologous RecombinationEfficiencyDouble strand BreaksEndonucleasesNonhomologous End JoiningNHEJHomology directed RepairHDR

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0:00

Overview

0:45

Principles of Genome Editing

4:10

Generalized CRISPR Protocol

6:11

Applications

8:03

Summary

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