ArticlePlant physiology2015
A Universal Positive-Negative Selection System for Gene Targeting in Plants Combining an Antibiotic Resistance Gene and Its Antisense RNA.
Article in Plant physiology, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 11 papers.
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Who cites it
11 citing papers in PubMed, 22 citations in OpenAlex.
- CRISPR/Cas-mediated genome editing: playing a versatile role in mitigating the challenges of sustainable rice improvement.3 Biotech · 2025Review
- Precise genetic engineering withPlant biotechnology (Tokyo, Japan) · 2023Article
- A piggyBac-mediated transgenesis system for the temporary expression of CRISPR/Cas9 in rice.Plant biotechnology journal · 2021Article
- CRISPR/Cas-based precision genome editing via microhomology-mediated end joining.Plant biotechnology journal · 2021Review
- Genome editing in rice.Rice (New York, N.Y.) · 2020Article
- Precise Genome Editing in miRNA Target Site via Gene Targeting and Subsequent Single-Strand-Annealing-Mediated Excision of the Marker Gene in Plants.Frontiers in genome editing · 2020Article
- Challenges and Perspectives in Homology-Directed Gene Targeting in Monocot Plants.Rice (New York, N.Y.) · 2019Review
- Progress of targeted genome modification approaches in higher plants.Plant cell reports · 2016Review
- Homology-based double-strand break-induced genome engineering in plants.Plant cell reports · 2016Review
- Precise Genome Modification via Sequence-Specific Nucleases-Mediated Gene Targeting for Crop Improvement.Frontiers in plant science · 2016Review
- Gene targeting and transgene stacking using intra genomic homologous recombination in plants.Plant methods · 2016Review
Corrections and comments
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Authors and funding
5 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Gene targeting (GT) is a useful technology for accurate genome engineering in plants. A reproducible approach based on a positive-negative selection system using hygromycin resistance and the diphtheria toxin A subunit gene as positive and negative selection markers, respectively, is now available. However, to date, this selection system has been applied exclusively in rice (Oryza sativa). To establish a universally applicable positive-negative GT system in plants, we designed a selection system using a combination of neomycin phosphotransferaseII (nptII) and an antisense nptII construct. The concomitant transcription of both sense and antisense nptII suppresses significantly the level of expression of the sense nptII gene, and transgenic calli and plants become sensitive to the antibiotic geneticin. In addition, we were able to utilize the sense nptII gene as a positive selection marker and the antisense nptII construct as a negative selection marker for knockout of the endogenous rice genes Waxy and 33-kD globulin through GT, although negative selection with this system is relatively less efficient compared with diphtheria toxin A subunit. The approach developed here, with some additional improvements, could be applied as a universal selection system for the enrichment of GT cells in several plant species.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.