Matrix attachment regions as a tool to influence plant transgene expression

Dolgova, Anna; Dolgov, Sergey

doi:10.1007/s13205-019-1709-5

Cited by 12 publications

(3 citation statements)

References 71 publications

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“…One likely reason could be that the variability in transgene expression is influenced not only by the copy number of transgene within the integration site but also by the chromatin status (heterochromatic or euchromatic). Additionally, transgene expression could be affected by nearby regulatory sequences, such as enhancers or silencers (Dolgova and Dolgov, 2019). Although the results of the sqRT-PCR analysis showed that every fluorescent gene was transcribed into RNA (Fig.…”

Section: Resultsmentioning

confidence: 99%

Strong Fluorescence Expression of ZsGreen1 in Petunia Flowers by Agrobacterium tumefaciens–mediated Transformation

Cho¹,

Kim²,

Alvarez³

et al. 2019

J. Amer. Soc. Hort. Sci.

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Fluorescent proteins (FT) have become essential, biological research tools. Many novel genes have been cloned from a variety of species and modified for effective, stable, and strong expression in transgenic organisms. Although there are many applications, FT expression has been employed most commonly at the cellular level in plants. To investigate FT expression at the whole-plant level, particularly in flowers, petunia ‘Mitchell Diploid’ [MD (Petunia ×hybrida)] was genetically transformed with seven genes encoding FTs: DsRed2, E2Crimson, TurboRFP, ZsGreen1, ZsYellow1, rpulFKz1, or aeCP597. Each gene was cloned into a pHK-DEST-OE vector harboring constitutive figwort mosaic virus 35S promoter and NOS-terminator. These plasmids were individually introduced into the genome of MD by Agrobacterium tumefaciens–mediated transformation. Shoot regeneration efficiency from the cocultured explants ranged from 8.3% to 20.3%. Various intensities of red, green, and yellow fluorescence were detected from TurboRFP, ZsGreen1, and ZsYellow1-transgenic flowers, respectively, under ultraviolet light for specific excitation and emission filters. More than 70% of plants established from the regenerated shoots were confirmed as transgenic plants. Transgenic ZsGreen1 petunia generated strong, green fluorescence in all flower organs of T0 plants including petals, stigmas, styles, anthers, and filaments. Most of the chromophores were localized to the cytoplasm but also went into the nuclei of petal cells. There was a positive linear relationship (R2 = 0.88) between the transgene expression levels and the relative fluorescent intensities of the ZsGreen1-transgenic flowers. No fluorescence was detected from the flowers of DsRed2-, E2Crimson-, rpulFKz1-, or aeCP597-transgenic petunias even though their gene transcripts were confirmed through semiquantitative reverse transcriptase-polymerase chain reaction. T1 generation ZsGreen1 plants showed green fluorescence emission from the cotyledons, hypocotyls, and radicles, which indicated stable FT expression was heritable. Four homozygous T2 inbred lines were finally selected. Throughout this study, we demonstrated that ZsGreen1 was most suitable for generating visible fluorescence in MD flowers among the seven genes tested. Thus, ZsGreen1 may have excellent potential for better utility as a sensitive selectable marker.

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Section: Resultsmentioning

confidence: 99%

Strong Fluorescence Expression of ZsGreen1 in Petunia Flowers by Agrobacterium tumefaciens–mediated Transformation

Cho¹,

Kim²,

Alvarez³

et al. 2019

J. Amer. Soc. Hort. Sci.

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“…Nuclear matrix attachment regions (MARs) are A/T-rich DNA sequences of approximately 400 bp found at the borders of nontranscribed genes that mediate the structural organization of chromatin within the nucleus (Dolgova and Dolgov, 2019). MAR sequences have been used to flank MCs to reduce transgene silencing, improve expression stability (Zhang et al, 2004;, and enhance transformation efficiency (Petersen et al, 2002).…”

Section: Binary and Alternative Vectorsmentioning

confidence: 99%

Insights Into Genetic and Molecular Elements for Transgenic Crop Development

et al. 2020

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Climate change and the exploration of new areas of cultivation have impacted the yields of several economically important crops worldwide. Both conventional plant breeding based on planned crosses between parents with specific traits and genetic engineering to develop new biotechnological tools (NBTs) have allowed the development of elite cultivars with new features of agronomic interest. The use of these NBTs in the search for agricultural solutions has gained prominence in recent years due to their rapid generation of elite cultivars that meet the needs of crop producers, and the efficiency of these NBTs is closely related to the optimization or best use of their elements. Currently, several genetic engineering techniques are used in synthetic biotechnology to successfully improve desirable traits or remove undesirable traits in crops. However, the features, drawbacks, and advantages of each technique are still not well understood, and thus, these methods have not been fully exploited. Here, we provide a brief overview of the plant genetic engineering platforms that have been used for proof of concept and agronomic trait improvement, review the major elements and processes of synthetic biotechnology, and, finally, present the major NBTs used to improve agronomic traits in socioeconomically important crops.

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“…MARs are often correlated with chromatin features like, AT richness, the origin of replication, etc., and are also hotspots for transposable element insertions (6). Many studies have been carried out determining their role in the maintenance of chromatin architecture, stabilization and regulation of gene expression (7)(8)(9) and their in uence on gene silencing mechanisms (10)(11)(12).…”

Section: Introductionmentioning

confidence: 99%

Dynamics of Nuclear Matrix Attachment Regions During 5th Instar Posterior Silk Gland Development in Bombyx Mori

Chunduri

Rajan

Lima

et al. 2022

Preprint

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Chromatin architecture is critical for gene expression during development. Matrix attachment regions (MARs) control and regulate chromatin dynamics. The position of MARs in the genome determines the expression of genes in the organism. In this study, we set out to elucidate how MARs temporally regulate the expression of the fibroin heavy chain (FIBH) gene during development. We addressed this by identifying MARs and studying their distribution and differentiation, in the posterior silk glands of Bombyx mori during 5th instar development. Of the MARs identified on three different days, 7.15% MARs were common to all three, whereas, 1.41%, 19.27% and 52.47% MARs were unique to day 1, day 5, and day 7, respectively highlighting the dynamic nature of the matrix associated DNA. Further, significant changes in the MARs in the vicinity of the FIBH gene were found during different days of 5th instar development which implied their role in the regulation and expression of the FIBH gene.

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Matrix attachment regions as a tool to influence plant transgene expression

Cited by 12 publications

References 71 publications

Strong Fluorescence Expression of ZsGreen1 in Petunia Flowers by Agrobacterium tumefaciens–mediated Transformation

Strong Fluorescence Expression of ZsGreen1 in Petunia Flowers by Agrobacterium tumefaciens–mediated Transformation

Insights Into Genetic and Molecular Elements for Transgenic Crop Development

Dynamics of Nuclear Matrix Attachment Regions During 5th Instar Posterior Silk Gland Development in Bombyx Mori

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