A self‐stabilizing <i>Ac</i> derivative and its potential for transposon tagging

Schmitz, Gregor; Theres, Klaus

doi:10.1046/j.1365-313x.1994.6050781.x

Cited by 12 publications

(4 citation statements)

References 24 publications

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“…This scenario is further supported by the family-specific detection of disproportionately more unique insertions onto chromosomes also harboring a common insertion (Table II). The implications of these data are that a given Ds element insertion site could be effectively utilized as a new launchpad for future insertions, simply by reintroducing the transposase gene through cross pollination of Ds-only plants with Aconly plants; the transactivation system described by Schmitz and Theres (1994) effectively demonstrates this model. Transposition of cis-acting elements to the entire genome has been demonstrated in rice (Kumar et al, 2005;Qu et al, 2008).…”

Section: Discussionmentioning

confidence: 99%

An ActiveAc/DsTransposon System for Activation Tagging in Tomato Cultivar M82 Using Clonal Propagation

et al. 2013

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Tomato (Solanum lycopersicum) is a model organism for Solanaceae in both molecular and agronomic research. This project utilized Agrobacterium tumefaciens transformation and the transposon-tagging construct Activator (Ac)/Dissociator (Ds)-ATag-Bar_gosGFP to produce activation-tagged and knockout mutants in the processing tomato cultivar M82. The construct carried hygromycin resistance (hyg), green fluorescent protein (GFP), and the transposase (TPase) of maize (Zea mays) Activator major transcript X054214.1 on the stable Ac element, along with a 35S enhancer tetramer and glufosinate herbicide resistance (BAR) on the mobile Ds-ATag element. An in vitro propagation strategy was used to produce a population of 25 T0 plants from a single transformed plant regenerated in tissue culture. A T1 population of 11,000 selfed and cv M82 backcrossed progeny was produced from the functional T0 line. This population was screened using glufosinate herbicide, hygromycin leaf painting, and multiplex polymerase chain reaction (PCR). Insertion sites of transposed Ds-ATag elements were identified through thermal asymmetric interlaced PCR, and resulting product sequences were aligned to the recently published tomato genome. A population of 509 independent, Ds-only transposant lines spanning all 12 tomato chromosomes has been developed. Insertion site analysis demonstrated that more than 80% of these lines harbored Ds insertions conducive to activation tagging. The capacity of the Ds-ATag element to alter transcription was verified by quantitative real-time reverse transcription-PCR in two mutant lines. The transposon-tagged lines have been immortalized in seed stocks and can be accessed through an online database, providing a unique resource for tomato breeding and analysis of gene function in the background of a commercial tomato cultivar.

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Section: Discussionmentioning

confidence: 99%

An ActiveAc/DsTransposon System for Activation Tagging in Tomato Cultivar M82 Using Clonal Propagation

et al. 2013

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“…Various modifications were made to insertional mutagenesis strategies and included, for example, a way to stabilize Ac once it had transposed to a different site (Schmitz and Theres, 1994). This entailed the use of a mutated Ac element (Ds303) containing transposase that lacked a promoter, so the initial transposition required an adjacent promoter, contained in the T-DNA, to initiate movement.…”

Section: Transposon-based Taggingmentioning

confidence: 99%

“…This entailed the use of a mutated Ac element (Ds303) containing transposase that lacked a promoter, so the initial transposition required an adjacent promoter, contained in the T-DNA, to initiate movement. It was demonstrated that A. tumefaciensgenerated transgenic tomato harboring this construct fostered one transposition before Ds303 became stable (nonautonomous; Schmitz and Theres, 1994). A gainof-function transposon mutagenesis approach used a similar tactic incorporating an engineered element (DsAT) that could transpose once, before becoming stable (Suzuki et al, 2001).…”

Section: Transposon-based Taggingmentioning

confidence: 99%

Genome Editing of Plants

Songstad

Petolino

Voytas

et al. 2017

Critical Reviews in Plant Sciences

111

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Genome editing in organisms via random mutagenesis is a naturally occurring phenomenon. As a technology, genome editing has evolved from the use of chemical and physical mutagenic agents capable of altering DNA sequences to biological tools such as designed sequence-specific nucleases (SSN) to produce knockout (KO) or knock-in (KI) edits and Oligonucleotide Directed Mutagenesis (ODM) where specific nucleotide changes are made in a directed manner resulting in custom single nucleotide polymorphisms (SNPs). Cibus' SU Canola TM , which the US Department of Agriculture (USDA) views as non-genetically modified (non-GM), is Cibus' first commercial product arising from plant genome editing and had its test launch in 2014. Regulatory aspects of the various genome editing tools will be discussed.

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“…The transposon sometimes undergoes further transposition without induction, possibly because of endogenous stimuli. Schmitz and Theres (1994) constructed a self-stabilizing Ac derivative whose 3 0 end located between the transposase gene and its promoter. The construct was improved for obtaining gain-of-function mutations (Suzuki et al 2001).…”

Section: Introductionmentioning

confidence: 99%

A one-time inducible transposon for creating knockout mutants

Lin

Huang

et al. 2008

Mol Breeding

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The maize transposon Ac can move to a new location within the genome to create knockout mutants in transgenic plants. In rice, Ac transposon is very active but sometimes undergoes further transposition and leaves an empty mutated gene. Therefore, we developed a one-time transposon system by locating one end of the transposon in the intron of the Ac transposase gene, which is under the control of the inducible promoter (PR-1a). Treatment with salicylic acid induced transposition of this transposon, COYA, leading to transposase gene breakage in exons. The progeny plants inheriting the transposition events become stable knockout mutants, because no functional transposase could be yielded. The behavior of COYA was analyzed in single-copy transgenic rice plants. We determined the expression of the modified transposase gene and its ability to trigger transposition events in transgenic rice plants. The COYA element thus exhibits potential for development of an inducible transposon system suitable for gene isolation in heterologous plant species.

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A self‐stabilizing Ac derivative and its potential for transposon tagging

Cited by 12 publications

References 24 publications

An ActiveAc/DsTransposon System for Activation Tagging in Tomato Cultivar M82 Using Clonal Propagation

An ActiveAc/DsTransposon System for Activation Tagging in Tomato Cultivar M82 Using Clonal Propagation

Genome Editing of Plants

A one-time inducible transposon for creating knockout mutants

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