Abstract
Plant-based systems are safe alternatives to the current platforms for the production of biologically active therapeutic proteins. However, plant-based expression systems face certain major challenges, including the relatively low productivity and the generation of target proteins in biologically active forms. The use of plant virus-based expression systems has been shown to enhance yields, but further improvement is still required to lower the production cost. In this study, various strategies were employed to increase the yields of an important therapeutic protein, human interferon gamma (IFNγ), in Nicotiana benthamiana through modifications of expression vectors based on potexviruses. Among these, the vector based on a coat protein (CP)-deficient Bamboo mosaic virus (BaMV), pKB△CHis, was shown to exhibit the highest expression level for the unmodified IFNγ. Truncation of the N-terminal signal peptide of IFN (designated mIFNγ) resulted in a nearly seven-fold increase in yield. Co-expression of a silencing suppressor protein by replacing the coding sequence of BaMV movement protein with that of P19 led to a 40% increase in mIFNγ accumulation. The fusion of endoplasmic reticulum (ER) retention signal with mIFNγ significantly enhanced the accumulation ratio of biologically active dimeric mIFNγ to 87% relative to the non-active monomeric form. The construct pKB19mIFNγER, employing the combination of all the above enhancement strategies, gave the highest level of protein accumulation, up to 119 ± 0.8 μg/g fresh weight, accounting for 2.5% of total soluble protein (TSP) content. These findings advocate the application of the modified BaMV-based vector as a platform for high-level expression of therapeutic protein in N. benthamiana.
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Claims 6
Claim · model-written summary
The pKB△CHis BaMV vector gave the highest expression level of unmodified IFNγ among the modified vectors.
- Taxon
- Nicotiana benthamiana
Nicotiana benthamianat0 - Treatment
- pKB△CHise1
- Gene
- IFNγe1
- Outcome
- expression levele1
- Direction
- higheste1
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“Nicotiana benthamiana”
e1“the vector based on a coat protein (CP)-deficient Bamboo mosaic virus (BaMV), pKB△CHis, was shown to exhibit the highest expression level for the unmodified IFNγ”
Claim · model-written summary
Truncating the N-terminal signal peptide of IFN (mIFNγ) gave a nearly seven-fold increase in yield.
- Taxon
- Nicotiana benthamianaNicotiana benthamianat0
- Treatment
- Truncation of the N-terminal signal peptidee2
- Gene
- mIFNγe2
- Outcome
- yielde2
- Direction
- increasee2
- Value
- nearly seven-folde2
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“Nicotiana benthamiana”
e2“Truncation of the N-terminal signal peptide of IFN (designated mIFNγ) resulted in a nearly seven-fold increase in yield”
Claim · model-written summary
Co-expressing the silencing suppressor P19 in place of the BaMV movement protein gave a 40% increase in mIFNγ accumulation.
- Taxon
- Nicotiana benthamianaNicotiana benthamianat0
- Treatment
- Co-expression of a silencing suppressor protein by replacing the coding sequence of BaMV movement protein with that of P19e3
- Gene
- P19e3
- Outcome
- mIFNγ accumulatione3
- Direction
- increasee3
- Value
- 40%e3
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“Nicotiana benthamiana”
e3“Co-expression of a silencing suppressor protein by replacing the coding sequence of BaMV movement protein with that of P19 led to a 40% increase in mIFNγ accumulation”
Claim · model-written summary
Fusing an ER retention signal to mIFNγ significantly enhanced the ratio of biologically active dimeric mIFNγ to 87%, relative to the non-active monomeric form.
- Taxon
- Nicotiana benthamianaNicotiana benthamianat0
- Treatment
- fusion of endoplasmic reticulum (ER) retention signal with mIFNγe4
- Gene
- mIFNγe4
- Outcome
- accumulation ratio of biologically active dimeric mIFNγe4
- Direction
- enhancede4
- Value
- 87%e4
- Comparator
- non-active monomeric forme4
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“Nicotiana benthamiana”
e4“The fusion of endoplasmic reticulum (ER) retention signal with mIFNγ significantly enhanced the accumulation ratio of biologically active dimeric mIFNγ to 87% relative to the non-active monomeric form”
Claim · model-written summary
The pKB19mIFNγER construct, combining all enhancement strategies, gave the highest protein accumulation, up to 119 ± 0.8 μg/g fresh weight.
- Taxon
- Nicotiana benthamianaNicotiana benthamianat0
- Treatment
- pKB19mIFNγERe5
- Outcome
- protein accumulatione5
- Direction
- higheste5
- Value
- 119 ± 0.8 μg/g fresh weighte5
- Qualifier
- accounting for 2.5% of total soluble proteine5
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“Nicotiana benthamiana”
e5“The construct pKB19mIFNγER, employing the combination of all the above enhancement strategies, gave the highest level of protein accumulation, up to 119 ± 0.8 μg/g fresh weight, accounting for 2.5% of total soluble protein (TSP) content”
Claim · model-written summary
The authors advocate the modified BaMV-based vector as a platform for high-level expression of therapeutic protein in N. benthamiana.
- Taxon
- N. benthamianaNicotiana benthamianae6
- Outcome
- application of the modified BaMV-based vector as a platforme6
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e6“These findings advocate the application of the modified BaMV-based vector as a platform for high-level expression of therapeutic protein in N. benthamiana”
The same questions in other papers
loss of function · molecular: metabolites · Nicotiana benthamianaPositive and negative results
9 papers in the same direction as this paper · 25 in the opposite direction
- negative
- negative
- no effect
31 papers more in this group
See all 35 papers →Claims are grouped by organism, treatment and what was measured or targeted. A paper counts as positive when any of its claims in the group is positive.