Unexpected failure rates for modular assembly of engineered zinc fingers

By C. L. Ramirez, Jonathan E. Foley, David A. Wright, Felix Müller-Lerch, S. H. Rahman, T. Cornu, Ronnie J. Winfrey, Jeffry D. Sander, Fengli Fu, J. Townsend and 3 more. Nature Methods, 2008

447 citations recorded by the source corpus.

Subfields: Electrical and Electronic Engineering, Molecular Biology

View Paper: https://www.semanticscholar.org/paper/9960963dd523f48c86e328fea09f944a9902c091

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Citations in the corpus

• Genome editing with engineered zinc finger nucleases (2010, background): “Although such interactions can enable more selective binding, they can also complicate efforts to produce new ZFPs through modular desig”
• Rapid “open-source” engineering of customized zinc-finger nucleases for highly efficient gene modification (2008, unknown): “Although easy to perform, modular assembly has an efficacy rate for making functional ZFN pairs that is less than 6% (Ramirez et al., 2008) and can yield ZFNs with low activities and/or high toxicities (Cornu et al., 2008; Pruett- Miller et al., 2008).”
• Genome Engineering With Zinc-Finger Nucleases (2011, background): “In Review 777 fact, a substantial proportion of ZFN pairs fail, whether they are produced by design or selection (Ramirez et al. 2008; Joung et al. 2010; Kim et al. 2010).”
• Genome Engineering of Crops with Designer Nucleases (2012, background): “…build ZFNs by standard restriction digestion-based subcloning techniques; however, many of the ZFNs made by this approach are not highly active (Ramirez et al., 2008). h e reason is that modular assembly treats each i nger as an independent unit, failing to account for context-dependent inl…”
• The CRISPR/Cas9 system for plant genome editing and beyond. (2015, background): “Commercially available ZFNs generally perform better than those designed using publicly available resources but they are much more expensive (Ramirez et al., 2008).”
• Delivering CRISPR: a review of the challenges and approaches (2018, background): “Challenges with ZFNs include design and engineering of the ZFP for high-affinity binding of the desired sequence, which can prove non-trivial (Ramirez et al., 2008).”
• TALENs: a widely applicable technology for targeted genome editing (2012, unknown): “Modular assembly of individual pre-selected zinc finger domains provides a simple and rapid method for creating customized arrays 88 but the success rate of this strategy has been reported to be low 89 .”