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Cited by 486

The DOI matches its Crossref record; 30 of the 30 references it cites were found in a registry or an index.

Transcription Activator-Like Effector Nucleases Enable Efficient Plant Genome Engineering

Open access
Zhang, Yong, Zhang, Feng, Li, Xiaohong, Baller, Joshua A., Qi, Yiping, Starker, Colby G., Bogdanove, Adam J., Voytas, Daniel F.·2012·Plant Physiology

Abstract

The ability to precisely engineer plant genomes offers much potential for advancing basic and applied plant biology. Here, we describe methods for the targeted modification of plant genomes using transcription activator-like effector nucleases (TALENs). Methods were optimized using tobacco (Nicotiana tabacum) protoplasts and TALENs targeting the acetolactate synthase (ALS) gene. Optimal TALEN scaffolds were identified using a protoplast-based single-strand annealing assay in which TALEN cleavage creates a functional yellow fluorescent protein gene, enabling quantification of TALEN activity by flow cytometry. Single-strand annealing activity data for TALENs with different scaffolds correlated highly with their activity at endogenous targets, as measured by high-throughput DNA sequencing of polymerase chain reaction products encompassing the TALEN recognition sites. TALENs introduced targeted mutations in ALS in 30% of transformed cells, and the frequencies of targeted gene insertion approximated 14%. These efficiencies made it possible to recover genome modifications without selection or enrichment regimes: 32% of tobacco calli generated from protoplasts transformed with TALEN-encoding constructs had TALEN-induced mutations in ALS, and of 16 calli characterized in detail, all had mutations in one allele each of the duplicate ALS genes (SurA and SurB). In calli derived from cells treated with a TALEN and a 322-bp donor molecule differing by 6 bp from the ALS coding sequence, 4% showed evidence of targeted gene replacement. The optimized reagents implemented in plant protoplasts should be useful for targeted modification of cells from diverse plant species and using a variety of means for reagent delivery.

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Metadata

Authors
Zhang, Y., Zhang, F., Li, X., Baller, J. A., Qi, Y., Starker, C. G., Bogdanove, A. J., & Voytas, D. F.
Venue
Plant Physiology
Published
2012-11-02
Volume
161
Issue
1
Pages
20-27
ISSN
  • 0032-0889 (ISSN-L)
  • 1532-2548
OpenAlex
W2135008913
Cited by
486
References
30
FWCI
16.84
Type
journal article
Language
en
Open access
hybrid

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CoChat. (2026). Citation check: Transcription Activator-Like Effector Nucleases Enable Efficient Plant Genome Engineering. https://app.cochat.ai/papers/10.1104/pp.112.205179
The paper
Zhang, Y., Zhang, F., Li, X., Baller, J. A., Qi, Y., Starker, C. G., Bogdanove, A. J., & Voytas, D. F. (2012). Transcription Activator-Like Effector Nucleases Enable Efficient Plant Genome Engineering. Plant Physiology. https://doi.org/10.1104/pp.112.205179

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