Telomere‐Mediated Chromosomal Truncation for Generating Engineered Minichromosomes in Maize

Swyers, Nathan C.; Cody, Jon P.; McCaw, Morgan; Graham, Nathaniel D.; Zhao, Changzeng; Gaeta, Robert T.; Birchler, James A.

doi:10.1002/cppb.20031

Cited by 5 publications

(5 citation statements)

References 29 publications

(30 reference statements)

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“…The typical size range of a centromere satellite unit is 120 to 180 base pairs (bp), which is approximately the length of DNA required to encircle a nucleosome that provides the initial level of DNA packaging within chromatin. For example, the length of a monomer DNA satellite is 178 bp in Arabidopsis thaliana, 177 bp in radish ( Raphanus sativus ), and 156 bp in maize . However, satellite DNA repeat units as short as 20 bp have been found in Astragalus sinicus and as long as 5390 bp in potatoes .…”

Section: Plant Artificial Centromeresmentioning

confidence: 99%

“…For example, the length of a monomer DNA satellite is 178 bp in Arabidopsis thaliana, 13 177 bp in radish (Raphanus sativus), 14 and 156 bp in maize. 15 However, satellite DNA repeat units as short as 20 bp have been found in Astragalus sinicus 16 and as long as 5390 bp in potatoes. 17 Until now, the technical constraints in amplification and cloning processes make it challenging to obtain large tandem DNA repeats directly from plant genomes.…”

Section: ■ Plant Artificial Centromeresmentioning

confidence: 99%

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Plant Artificial Chromosomes: Construction and Transformation

Wang,

Lin,

et al. 2024

ACS Synth. Biol.

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With the decline of cultivated land and increase of the population in recent years, an agricultural revolution is urgently needed to produce more food to improve the living standards of humans. As one of the foundations of synthetic biology, artificial chromosomes hold great potential for advancing crop improvement. They offer opportunities to increase crop yield and quality, while enhancing crop resistance to disease. The progress made in plant artificial chromosome technology enables selective modification of existing chromosomes or the synthesis of new ones to improve crops and study gene function. However, current artificial chromosome technologies still face limitations, particularly in the synthesis of repeat sequences and the transformation of large DNA fragments. In this review, we will introduce the structure of plant centromeres, the construction of plant artificial chromosomes, and possible methods for transforming large fragments into plant cells.

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Section: Plant Artificial Centromeresmentioning

confidence: 99%

Section: ■ Plant Artificial Centromeresmentioning

confidence: 99%

Plant Artificial Chromosomes: Construction and Transformation

Wang,

Lin,

et al. 2024

ACS Synth. Biol.

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“…Moreover, the TMCT was faithfully transmitted to the next generation, and truncated chromosome can recombine with normal chromosome via Cre/lox or FLP/FRT recombination system, indicating the successful application of the TMCT platform ( Yu et al, 2007 ; Xu et al, 2012 ). Diverse length of Arabidopsis-type telomere repeat TTTAGGG were inserted into maize ( Yu et al, 2006 , 2007 ; Gaeta et al, 2011 ; Swyers et al, 2016 ), rice ( Oryza sativa ; Xu et al, 2012 ; Xu and Yu, 2016 ), Arabidopsis ( Nelson et al, 2011 ; Teo et al, 2011 ), barley ( Hordeum vulgare ; Kapusi et al, 2012 ), wheat ( Triticum aestivum ; Yuan et al, 2017 ), and Brassica napus ( Yan et al, 2017 ; Yin et al, 2021 ) by agrobacterium-mediated transformation or biotransformation, and TMCT were generated, which revealed that minichromosomes could be created in most of plant species by TMCT approach.…”

Section: Plant Artificial Chromosomesmentioning

confidence: 99%

Artificial chromosome technology and its potential application in plants

2022

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Plant genetic engineering and transgenic technology are powerful ways to study the function of genes and improve crop yield and quality in the past few years. However, only a few genes could be transformed by most available genetic engineering and transgenic technologies, so changes still need to be made to meet the demands for high throughput studies, such as investigating the whole genetic pathway of crop traits and avoiding undesirable genes simultaneously in the next generation. Plant artificial chromosome (PAC) technology provides a carrier which allows us to assemble multiple and specific genes to produce a variety of products by minichromosome. However, PAC technology also have limitations that may hinder its further development and application. In this review, we will introduce the current state of PACs technology from PACs formation, factors on PACs formation, problems and potential solutions of PACs and exogenous gene(s) integration.

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“…It contains 2.5 kb of Arabidopsis telomere sequence. Using the cloning protocol described (Swyers et al 2016), 2 kb of telomere sequence was placed into WY9PZPNOS 1.4, just inside of the Right Border sequence. Stbl 4 cells were used to stabilize the telomere repeat.…”

Section: Construct Designmentioning

confidence: 99%

“…Appendix 2 describes cloning of telomere DNA into a construct in detail. Positive transformants were screened using a colony hybridization and Southern Blot analysis (Swyers et al 2016) (Figure 6).…”

Section: Construct Designmentioning

confidence: 99%

Development of an amenable system for site-specific addition to a maize chromosome

Swyers¹

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Currently, transgenic maize is produced by random integration of a transgenes into the plant. This works for single genes, but not as well for multiple traits. Identifying plants that contain several transgenes becomes a very difficult task. Gene stacking at a single location in the genome would make combining multiple transgenes into plants a simpler process. This project focused on the development of a system would allow for transgenes to be sequentially added to a specific site in the maize genome. The system utilizes two recombinases, Cre recombinase and _C31 Integrase, to remove a selectable marker and to integrate transgenes. An initial construct containing a selectable marker, flanked by LoxP sites, which are acted upon by Cre recombinase, and an attP site, were transformed. The selectable marker was then removed from the integrated transgene by exposure to Cre recombinase. Two amendment constructs enable modification of the integrated construct by utilizing complementary attP and attB sites, which are acted upon by _C31 Integrase. The amendment constructs contain cargo and a promoterless selectable marker which, upon successful recombination with the target site, restores expression of the selectable marker. Successful demonstration of this system simplifies generation of multi-transgene plants, and the assembly of multi-gene pathways in plants.

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Telomere‐Mediated Chromosomal Truncation for Generating Engineered Minichromosomes in Maize

Cited by 5 publications

References 29 publications

Plant Artificial Chromosomes: Construction and Transformation

Plant Artificial Chromosomes: Construction and Transformation

Artificial chromosome technology and its potential application in plants

Development of an amenable system for site-specific addition to a maize chromosome

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