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JoVE Journal
Genetics

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A Standard Methodology to Examine On-site Mutagenicity As a Function of Point Mutation Repair Catalyzed by CRISPR/Cas9 and SsODN in Human Cells
 

A Standard Methodology to Examine On-site Mutagenicity As a Function of Point Mutation Repair Catalyzed by CRISPR/Cas9 and SsODN in Human Cells

Article DOI: 10.3791/56195 10:08 min August 25th, 2017
August 25th, 2017

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Summary

This protocol outlines the workflow of a CRISPR/Cas9-based gene editing system for the repair of point mutations in mammalian cells. Here, we use a combinatorial approach to gene editing with a detailed follow-on experimental strategy for measuring indel formation at the target site—in essence, analyzing onsite mutagenesis.

Transcript

Tags

Standard Methodology Examine On-site Mutagenicity Point Mutation Repair CRISPR/Cas9 SsODN Human Cells Genetic Heterogeneity Gene Editing Field Onsite Mutagenesis Analysis Homology Directed Repair Gene Correction Single Stranded Oligonucleotides HTT11619 Cells T-175 Flask Dulbecco's Phosphate Buffered Saline (PBS) Trypsin Complete Medium Cell Clumps 15 Milliliter Conical Tube Trypan Blue
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