Engineered RNA-binding protein for transgene activation in non-green plastids

Research output: Contribution to journalLetter

4 Citations (Scopus)

Abstract

Non-green plastids are desirable for the expression of recombinant proteins in edible plant parts to enhance the nutritional value of tubers or fruits, or to deliver pharmaceuticals. However, plastid transgenes are expressed at extremely low levels in the amyloplasts of storage organs such as tubers 1–3 . Here, we report a regulatory system comprising a variant of the maize RNA-binding protein PPR10 and a cognate binding site upstream of a plastid transgene that encodes green fluorescent protein (GFP). The binding site is not recognized by the resident potato PPR10 protein, restricting GFP protein accumulation to low levels in leaves. When the PPR10 variant is expressed from the tuber-specific patatin promoter, GFP accumulates up to 1.3% of the total soluble protein, a 60-fold increase compared with previous studies 2 (0.02%). This regulatory system enables an increase in transgene expression in non-photosynthetic plastids without interfering with chloroplast gene expression in leaves.

Original languageEnglish (US)
Pages (from-to)486-490
Number of pages5
JournalNature Plants
Volume5
Issue number5
DOIs
StatePublished - May 1 2019

Fingerprint

RNA-binding proteins
transgenes
plastids
green fluorescent protein
tubers
binding sites
patatin
potato protein
amyloplasts
storage organs
recombinant proteins
plant anatomy
leaves
proteins
chloroplasts
nutritive value
promoter regions
drugs
gene expression
fruits

All Science Journal Classification (ASJC) codes

  • Plant Science

Cite this

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abstract = "Non-green plastids are desirable for the expression of recombinant proteins in edible plant parts to enhance the nutritional value of tubers or fruits, or to deliver pharmaceuticals. However, plastid transgenes are expressed at extremely low levels in the amyloplasts of storage organs such as tubers 1–3 . Here, we report a regulatory system comprising a variant of the maize RNA-binding protein PPR10 and a cognate binding site upstream of a plastid transgene that encodes green fluorescent protein (GFP). The binding site is not recognized by the resident potato PPR10 protein, restricting GFP protein accumulation to low levels in leaves. When the PPR10 variant is expressed from the tuber-specific patatin promoter, GFP accumulates up to 1.3{\%} of the total soluble protein, a 60-fold increase compared with previous studies 2 (0.02{\%}). This regulatory system enables an increase in transgene expression in non-photosynthetic plastids without interfering with chloroplast gene expression in leaves.",
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Engineered RNA-binding protein for transgene activation in non-green plastids. / Maliga, Pal.

In: Nature Plants, Vol. 5, No. 5, 01.05.2019, p. 486-490.

Research output: Contribution to journalLetter

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