High frequency modification of plant genes using engineered zinc finger nucleases

By J. Townsend, David A. Wright, Ronnie J. Winfrey, Fengli Fu, Morgan L. Maeder, J. Joung, Daniel F. Voytas. Nature, 2009

731 citations recorded by the source corpus.

An efficient method for making directed DNA sequence modifications to plant genes (gene targeting) is at present lacking, thereby frustrating efforts to dissect plant gene function and engineer crop plants that better meet the world’s burgeoning need for food, fibre and fuel. Zinc-finger nucleases (ZFNs)—enzymes engineered to create DNA double-strand breaks at specific loci—are potent stimulators of gene targeting; for example, they can be used to precisely modify engineered reporter genes in plants. Here we demonstrate high-frequency ZFN-stimulated gene targeting at endogenous plant genes, namely the tobacco acetolactate synthase genes (ALS SuRA and SuRB), for which specific mutations are known to confer resistance to imidazolinone and sulphonylurea herbicides. Herbicide-resistance mutations were introduced into SuR loci by ZFN-mediated gene targeting at frequencies exceeding 2% of transformed cells for mutations as far as 1.3 kilobases from the ZFN cleavage site. More than 40% of recombinant plants had modifications in multiple SuR alleles. The observed high frequency of gene targeting indicates that it is now possible to efficiently make targeted sequence changes in endogenous plant genes.

Subfields: Molecular Biology, Plant Science

View Paper: https://www.semanticscholar.org/paper/b2288b84366e7097126a99178bd9e1ba51105a71

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Citations in the corpus

A TALE nuclease architecture for efficient genome editing (2011, background): “, a setting that does not necessarily predict function on endogenous genes in more complex genome”
ZiFiT (Zinc Finger Targeter): an updated zinc finger engineering tool (2010, methodology): “Although this method requires considerable effort, it reliably generates ZFPs that bind with high affinity and specificity to their intended target site (7,9,11,16).”
Targeted Mutagenesis, Precise Gene Editing, and Site-Specific Gene Insertion in Maize Using Cas9 and Guide RNA[OPEN] (2015, background): “The frequency of stable event recovery has been reported to be an indicator of cytotoxicity caused by off-target recognition for ZFNs in human cell lines and plants (Szczepek et al., 2007; Cornu et al., 2008; Townsend et al., 2009).”
Efficient design and assembly of custom TALEN and other TAL effector-based constructs for DNA targeting (2011, methodology): “Similarly, we previously created herbicide resistant tobacco plants by gene targeting with ZFNs that recognize and cleave the acetolactate synthase gene (24).”
TAL nucleases (TALNs): hybrid proteins composed of TAL effectors and FokI DNA-cleavage domain (2010, methodology): “The yeast strains YPH499 (MATa ura3-52 lys2-801_amber ade2-101_ochre trp1-D63 his3-D200 leu2-D1) and YPH500 (MATa ura3-52 lys2-801_amber ade2-101_ochre trp1-D63 his3-D200 leu2-D1) as well as expression vectors (pCP3, pCP4 and pCP5) were described and kindly provided by Dr Dan Voytas (34).”
Rewiring of the Fruit Metabolome in Tomato Breeding. (2018, background): “High throughput molecular breeding combined with precision genome editing has huge potential to accelerate crop improvement (Soyk et al., 2017; Townsend et al., 2009), and reduce or eliminate linkage drag.”
TALENs: a widely applicable technology for targeted genome editing (2012, neutral): “…38 , catfish 39 , sea urchin 40 , frog 41 , pig 42 , cattle 43 , cricket 8 , rabbit 44 , silkworm 45 , butterfly 46 , mouse 47, 48 , rat 49, 50 , soybean 51, 52 , thale cress 53, 54 , corn 55 , tobacco 56 , petunia 57 , hamster cells 58 , and human somatic 59, 60 and pluripotent stem cells 61–63 .”
RNA-guided genome editing in plants using a CRISPR-Cas system. (2013, methodology): “Both ZFN and TALEN have been used in plant gene targeting and genome editing (Shukla et al., 2009; Townsend et al., 2009; Mahfouz et al., 2011; Weinthal et al., 2013; Zhang et al., 2013).”
Genome Engineering With Zinc-Finger Nucleases (2011, background): “Direct DNA transformation (Wright et al. 2005; Cai et al. 2009; Shukla et al. 2009; Townsend et al. 2009) and viral delivery (Marton et al. 2010) have also succeeded in plants.”
A CRISPR/Cas9 Toolbox for Multiplexed Plant Genome Editing and Transcriptional Regulation1[OPEN] (2015, background): “…to successfully target endogenous plant genes, creatingmutations, gene replacements, deletions, or inversions in Arabidopsis (Arabidopsis thaliana), tobacco (Nicotiana benthamiana), and soybean (Glycine max; Townsend et al., 2009; Zhang et al., 2010; Curtin et al., 2011; Qi et al., 2013b, 2014).”