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Updated: Jul 17, 2025

Genome Editing in Mammalian Cell Lines using CRISPR-Cas
Published on: April 11, 2019
PCR cloning Intermediated Gibson assembly (PIG) for Constructing DNA Repair Templates in CRISPR-Cas9 Based Gene
Dongsheng Han1, Scott Churcher1, Jared T Nordman1
1Department of Biological Sciences, Vanderbilt University, Nashville, Tennessee, United States.
Abstract:
The CRISPR-Cas9 gene-editing system has revolutionized genome engineering, allowing precise modifications to be made in a wide range of organisms. One significant challenge associated with CRISPR-Cas9 mediated gene editing is the construction of DNA repair templates containing homology arms, a screenable marker and a tag sequence of interest. Here, we present an efficient, two-step strategy to generate DNA repair templates in approximately one week, facilitating rapid and precise genome engineering applications.
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