Abstract
Clustered regularly interspaced short palindromic repeat (CRISPR) and other gene editing technologies such as zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs) show great promises for research and therapeutic applications. One major concern is the off-target effects generated by these nucleases at unintended genomic sequences. In silico methods are usually used for off-target site prediction. However, based on currently available algorithms, the predicted cleavage activity at these potential off-target sites does not always reflect the true cleavage in vivo. Here we present an unbiased screening protocol using integration-defective lentiviral vector (IDLV) and deep sequencing to map the off-target sites generated by gene editing tools.
Key words
- CRISPR/Cas9
- Genome editing
- IDLV
- Off-target activity
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Acknowledgments
This work is supported by a grant RB3-02161 from California Institute of Regenerative Medicine (J.K.Y.) and a grant from Nesvig Foundation.
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Wang, X., Wu, Y., Yee, JK. (2021). Detection of CRISPR/Cas9-Generated Off-Target Effect by Integration-Defective Lentiviral Vector. In: Fulga, T.A., Knapp, D.J.H.F., Ferry, Q.R.V. (eds) CRISPR Guide RNA Design. Methods in Molecular Biology, vol 2162. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0687-2_14
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DOI: https://doi.org/10.1007/978-1-0716-0687-2_14
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Publisher Name: Humana, New York, NY
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