2A-linked bi-, tri-, and quad-cistrons for the stepwise biosynthesis of β-carotene, zeaxanthin, and ketocarotenoids in rice endosperm

Ye Sol Jeong; Hyung Keun Ku; Young Joo Jung; Jae Kwang Kim; Kyoung Bok Lee; Ju Kon Kim; Sun Hyung Lim; Dongho Lee; Sun Hwa Ha

doi:10.1016/j.mec.2021.e00166

2A-linked bi-, tri-, and quad-cistrons for the stepwise biosynthesis of β-carotene, zeaxanthin, and ketocarotenoids in rice endosperm

Ye Sol Jeong, Hyung Keun Ku, Young Joo Jung, Jae Kwang Kim, Kyoung Bok Lee, Ju Kon Kim, Sun Hyung Lim, Dongho Lee, Sun Hwa Ha

Department of Biosystems and Biotechnology

Research output: Contribution to journal › Article › peer-review

4 Citations (Scopus)

Abstract

Foot-and-mouth disease virus (FMDV) 2A constructs have been successfully used for the production of “Golden Rice”, a β-carotene producing rice strain. However, to allay public fears and opposition to plants carrying a mammalian pathogenic viral sequence, 2A-like synthetic sequences from Thosea asigna virus and Infectious myonecrosis virus were used to coordinate the coexpression of carotenoid biosynthetic genes. Here, up to four carotenogenic genes encoding PSY, CRTI, BCH and BKT were concatenated and produced β-carotene, zeaxanthin, and ketocarotenoids (astaxanthin and adonixanthin) in transgenic rice seeds displaying color variation due to the difference in carotenoid content and composition.

Original language	English
Article number	e00166
Journal	Metabolic Engineering Communications
Volume	12
DOIs	https://doi.org/10.1016/j.mec.2021.e00166
Publication status	Published - 2021 Jun

Bibliographical note

Funding Information:
Protein levels were first examined to compare the cleavage efficiency among T2A, I2A, and F2A using all bicistronic lines of stPTAC and stPIAC with a representative line of previous stPAC including a F2A (Fig. 2c, top). In an immune-blot with seed proteins (40 μg), T2A exhibited a 1.2- and 2.4-fold higher “cleavage” efficiency compared with I2A2 and F2A respectively. Moreover, T2A and I2A2 did not generate the any un-cleaved product, unlike F2A, which produced PSY:F2A:TP-CRTI – this suggests improved “cleavage” activity with T2A and I2A2. Further, PSY:2A forms between the stPTAC and stPIAC lines were similar using 2A antibody (Fig. 2c, bottom; Fig. 2d, left). It supported the better function of T2A than I2A2 contributed to the translation of the second CRTI ORF (Fig. 2c, top).This work was supported by grants from the Next-Generation BioGreen 21 Program (PJ01368801) and the Next Generation New Plant Breed Technology Program (PJ01477202) funded by the Rural Development Administration.

Funding Information:
This work was supported by grants from the Next-Generation BioGreen 21 Program ( PJ01368801 ) and the Next Generation New Plant Breed Technology Program ( PJ01477202 ) funded by the Rural Development Administration .

Publisher Copyright:
© 2021 The Authors

Keywords

2A peptide
Carotenoid
Metabolic engineering
Polycistron
Rice

ASJC Scopus subject areas

Endocrinology, Diabetes and Metabolism
Biomedical Engineering

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/j.mec.2021.e00166

Cite this

@article{f3f0462608e54492b107ba675683f191,

title = "2A-linked bi-, tri-, and quad-cistrons for the stepwise biosynthesis of β-carotene, zeaxanthin, and ketocarotenoids in rice endosperm",

abstract = "Foot-and-mouth disease virus (FMDV) 2A constructs have been successfully used for the production of “Golden Rice”, a β-carotene producing rice strain. However, to allay public fears and opposition to plants carrying a mammalian pathogenic viral sequence, 2A-like synthetic sequences from Thosea asigna virus and Infectious myonecrosis virus were used to coordinate the coexpression of carotenoid biosynthetic genes. Here, up to four carotenogenic genes encoding PSY, CRTI, BCH and BKT were concatenated and produced β-carotene, zeaxanthin, and ketocarotenoids (astaxanthin and adonixanthin) in transgenic rice seeds displaying color variation due to the difference in carotenoid content and composition.",

keywords = "2A peptide, Carotenoid, Metabolic engineering, Polycistron, Rice",

author = "Jeong, {Ye Sol} and Ku, {Hyung Keun} and Jung, {Young Joo} and Kim, {Jae Kwang} and Lee, {Kyoung Bok} and Kim, {Ju Kon} and Lim, {Sun Hyung} and Dongho Lee and Ha, {Sun Hwa}",

note = "Funding Information: Protein levels were first examined to compare the cleavage efficiency among T2A, I2A, and F2A using all bicistronic lines of stPTAC and stPIAC with a representative line of previous stPAC including a F2A (Fig. 2c, top). In an immune-blot with seed proteins (40 μg), T2A exhibited a 1.2- and 2.4-fold higher “cleavage” efficiency compared with I2A2 and F2A respectively. Moreover, T2A and I2A2 did not generate the any un-cleaved product, unlike F2A, which produced PSY:F2A:TP-CRTI – this suggests improved “cleavage” activity with T2A and I2A2. Further, PSY:2A forms between the stPTAC and stPIAC lines were similar using 2A antibody (Fig. 2c, bottom; Fig. 2d, left). It supported the better function of T2A than I2A2 contributed to the translation of the second CRTI ORF (Fig. 2c, top).This work was supported by grants from the Next-Generation BioGreen 21 Program (PJ01368801) and the Next Generation New Plant Breed Technology Program (PJ01477202) funded by the Rural Development Administration. Funding Information: This work was supported by grants from the Next-Generation BioGreen 21 Program ( PJ01368801 ) and the Next Generation New Plant Breed Technology Program ( PJ01477202 ) funded by the Rural Development Administration . Publisher Copyright: {\textcopyright} 2021 The Authors",

year = "2021",

month = jun,

doi = "10.1016/j.mec.2021.e00166",

language = "English",

volume = "12",

journal = "Metabolic Engineering Communications",

issn = "2214-0301",

publisher = "Elsevier BV",

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T1 - 2A-linked bi-, tri-, and quad-cistrons for the stepwise biosynthesis of β-carotene, zeaxanthin, and ketocarotenoids in rice endosperm

AU - Jeong, Ye Sol

AU - Ku, Hyung Keun

AU - Jung, Young Joo

AU - Kim, Jae Kwang

AU - Lee, Kyoung Bok

AU - Kim, Ju Kon

AU - Lim, Sun Hyung

AU - Lee, Dongho

AU - Ha, Sun Hwa

N1 - Funding Information: Protein levels were first examined to compare the cleavage efficiency among T2A, I2A, and F2A using all bicistronic lines of stPTAC and stPIAC with a representative line of previous stPAC including a F2A (Fig. 2c, top). In an immune-blot with seed proteins (40 μg), T2A exhibited a 1.2- and 2.4-fold higher “cleavage” efficiency compared with I2A2 and F2A respectively. Moreover, T2A and I2A2 did not generate the any un-cleaved product, unlike F2A, which produced PSY:F2A:TP-CRTI – this suggests improved “cleavage” activity with T2A and I2A2. Further, PSY:2A forms between the stPTAC and stPIAC lines were similar using 2A antibody (Fig. 2c, bottom; Fig. 2d, left). It supported the better function of T2A than I2A2 contributed to the translation of the second CRTI ORF (Fig. 2c, top).This work was supported by grants from the Next-Generation BioGreen 21 Program (PJ01368801) and the Next Generation New Plant Breed Technology Program (PJ01477202) funded by the Rural Development Administration. Funding Information: This work was supported by grants from the Next-Generation BioGreen 21 Program ( PJ01368801 ) and the Next Generation New Plant Breed Technology Program ( PJ01477202 ) funded by the Rural Development Administration . Publisher Copyright: © 2021 The Authors

PY - 2021/6

Y1 - 2021/6

N2 - Foot-and-mouth disease virus (FMDV) 2A constructs have been successfully used for the production of “Golden Rice”, a β-carotene producing rice strain. However, to allay public fears and opposition to plants carrying a mammalian pathogenic viral sequence, 2A-like synthetic sequences from Thosea asigna virus and Infectious myonecrosis virus were used to coordinate the coexpression of carotenoid biosynthetic genes. Here, up to four carotenogenic genes encoding PSY, CRTI, BCH and BKT were concatenated and produced β-carotene, zeaxanthin, and ketocarotenoids (astaxanthin and adonixanthin) in transgenic rice seeds displaying color variation due to the difference in carotenoid content and composition.

AB - Foot-and-mouth disease virus (FMDV) 2A constructs have been successfully used for the production of “Golden Rice”, a β-carotene producing rice strain. However, to allay public fears and opposition to plants carrying a mammalian pathogenic viral sequence, 2A-like synthetic sequences from Thosea asigna virus and Infectious myonecrosis virus were used to coordinate the coexpression of carotenoid biosynthetic genes. Here, up to four carotenogenic genes encoding PSY, CRTI, BCH and BKT were concatenated and produced β-carotene, zeaxanthin, and ketocarotenoids (astaxanthin and adonixanthin) in transgenic rice seeds displaying color variation due to the difference in carotenoid content and composition.

KW - 2A peptide

KW - Carotenoid

KW - Metabolic engineering

KW - Polycistron

KW - Rice

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DO - 10.1016/j.mec.2021.e00166

M3 - Article

AN - SCOPUS:85101306628

SN - 2214-0301

VL - 12

JO - Metabolic Engineering Communications

JF - Metabolic Engineering Communications

M1 - e00166

ER -

2A-linked bi-, tri-, and quad-cistrons for the stepwise biosynthesis of β-carotene, zeaxanthin, and ketocarotenoids in rice endosperm

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

UN SDGs

Access to Document

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