Efficient deletion of LoxP-flanked selectable marker genes from the genome of transgenic pigs by an engineered Cre recombinase

Huang, Xiaoling; Zou, Xian; Xu, Zhiqian; Tang, Fei; Shi, Junsong; Zheng, Enqin; Liu, Dewu; Moisyadi, Stefan; Urschitz, Johann; Wu, Zhenfang; Li, Zicong

doi:10.1007/s11248-020-00200-3

“…to remove selectable markers in transformants. For example, the FLP-CRE system could remove the selectable marker gene by a further cross to induce Cre expression (Huang et al 2020;Zhao et al 2019). Using the FLP-CRE system, AtGASA6 could also be removed from transgenic progenies.…”

Section: Improvement Of the Atgasa6-assisted Selection Systemmentioning

confidence: 99%

A novel non-antibiotic selectable marker GASA6 for plant transformation

Luo

¹

,

Gu

²

,

Wang

³

et al. 2021

Plant Cell Tiss Organ Cult

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Selectable markers help the transformed cell/tissue to survive in an otherwise lethal exposure of an antibiotic or herbicide. Unfortunately, almost all the traditional selectable markers are antibiotic and herbicide resistance genes, which are controversial on human health concerns and environmental impact. Novel plant-derived, non-antibiotic, and non-herbicide selectable markers are urgently needed in plant transformation. Our previous work showed that the seedlings of overexpression Arabidopsis lines of AtGASA6 survived on medium with a high concentration of sugar, which leads to the hypothesis that AtGASA6 could be a selectable marker on media with high or low sugar content. In this study, leaf explants of AtGASA6 overexpression tobacco lines regenerated shoots on sugar-free shooting medium while those of wild type could not. Moreover, the seeds of AtGASA6 overexpression tobacco lines germinated and grew into normal seedlings on sugar-free MS medium while those of WT could not. Attractively, no developmental defects were observed in AtGASA6 transgenic progenies. Using AtGASA6 as a selectable marker, overexpression tobacco lines of GAI, which restrains plant size, were created on sugar-free media. The GAI overexpression lines had a smaller plant size than that of control. Considering its plant-derived and non-antibiotic nature, GASA6 is promising to be used as a selectable marker in plant transformation.

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“…In recent years, emerging techniques have been developed to remove selectable markers in transformants. For example, the FLP-CRE system could remove the selectable marker gene by a further cross to induce Cre expression (Huang et al 2020;Zhao et al 2019). Using the FLP-CRE system, AtGASA6 could also be removed from transgenic progenies.…”

Section: Improvement Of the Atgasa6-assisted Selection Systemmentioning

confidence: 99%

A novel non-antibiotic selectable marker GASA6 for plant transformation

Luo

¹

,

Gu

²

,

Wang

³

et al. 2021

Preprint

0

View full text Add to dashboard Cite

Selectable markers help the transformed cell/tissue to survive in an otherwise lethal exposure of an antibiotic or herbicide. Unfortunately, almost all the traditional selectable markers are antibiotic and herbicide resistance genes, which are controversial on human health concerns and environmental impact. Novel plant-derived, non-antibiotic, and non-herbicide selectable markers are urgently needed in plant transformation. Our previous work showed that the seedlings of overexpression Arabidopsis lines of AtGASA6 survived on medium with a high concentration of sugar, which leads to the hypothesis that AtGASA6 could be a selectable marker on media with high or low sugar content. In this study, leaf explants of AtGASA6 overexpression tobacco lines regenerated shoots on sugar-free shooting medium while those of wild type could not. Moreover, the seeds of AtGASA6 overexpression tobacco lines germinated and grew into normal seedlings on sugar-free MS medium while those of WT could not. Attractively, no developmental defects were observed in AtGASA6 transgenic progenies. Using AtGASA6 as a selectable marker, overexpression tobacco lines of GAI, which restrains plant size, were created on sugar-free media. The GAI overexpression lines had a smaller plant size than that of control. Considering its plant-derived and non-antibiotic nature, GASA6 is promising to be used as a selectable marker in plant transformation.

show abstract