Detection of Off-Target Effect in Genome Editing by Integration-Defective Lentiviral VectorSeptember 30, 2020
Guangzhou Medical University (China) and Beckman Research Institute (USA) presented an unbiased screening protocol integration-defective lentiviral vector (IDLV) and deep sequencing to map the off-target sites generated by gene editing tools. The details are published in the Methods in Molecular Biology book series.
Clustered regularly interspaced short palindromic repeat (CRISPR) and other gene editing technologies including zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs) exhibit wide usage for research and therapeutics. One of the major concerns about these technologies is the off-target effects generated by nucleases at unintended genomic sequences. To address this concern, in silico methods are applied to project possible off-target sites. However, the predicted cleavage activity at these potential off-target sites does not always reflect the true cleavage in vivo with the use of available algorithms. Thus, a new screening protocol was developed by Xiaoling Wang, Youjun Wu, and Jiing-Kuan Yee.
Read more about the protocol in CRISPR Guide RNA Design.
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