Chloroplast Cell-Free Systems from Different Plant Species as a Rapid Prototyping Platform DOI Creative Commons
Clemens V. Böhm, René Inckemann, Michael Burgis

et al.

ACS Synthetic Biology, Journal Year: 2024, Volume and Issue: 13(8), P. 2412 - 2424

Published: July 19, 2024

Climate change poses a significant threat to global agriculture, necessitating innovative solutions. Plant synthetic biology, particularly chloroplast engineering, holds promise as viable approach this challenge. Chloroplasts present variety of advantageous traits for genetic but the development tools and part characterization in these organelles is hindered by lengthy time scales required generate transplastomic organisms. To address challenges, we have established versatile protocol generating highly active chloroplast-based cell-free gene expression (CFE) systems derived from diverse range plant species, including wheat (monocot), spinach, poplar trees (dicots). We show that work with conventionally used T7 RNA polymerase well endogenous polymerases, allowing detailed prototyping regulatory sequences at both transcription translation levels. demonstrate platform promoters 5' 3' untranslated regions (UTRs) higher expression, analyze collection 23 5'UTRs, 10 3'UTRs, 6 promoters, assessed their spinach extracts, found consistency patterns, suggesting cross-species compatibility. Looking forward, our CFE open new avenues offering understanding developing engineered plants, which could help meet demands changing climate.

Language: Английский

Chloroplast Cell-Free Systems from Different Plant Species as a Rapid Prototyping Platform DOI Creative Commons
Clemens V. Böhm, René Inckemann, Michael Burgis

et al.

ACS Synthetic Biology, Journal Year: 2024, Volume and Issue: 13(8), P. 2412 - 2424

Published: July 19, 2024

Climate change poses a significant threat to global agriculture, necessitating innovative solutions. Plant synthetic biology, particularly chloroplast engineering, holds promise as viable approach this challenge. Chloroplasts present variety of advantageous traits for genetic but the development tools and part characterization in these organelles is hindered by lengthy time scales required generate transplastomic organisms. To address challenges, we have established versatile protocol generating highly active chloroplast-based cell-free gene expression (CFE) systems derived from diverse range plant species, including wheat (monocot), spinach, poplar trees (dicots). We show that work with conventionally used T7 RNA polymerase well endogenous polymerases, allowing detailed prototyping regulatory sequences at both transcription translation levels. demonstrate platform promoters 5' 3' untranslated regions (UTRs) higher expression, analyze collection 23 5'UTRs, 10 3'UTRs, 6 promoters, assessed their spinach extracts, found consistency patterns, suggesting cross-species compatibility. Looking forward, our CFE open new avenues offering understanding developing engineered plants, which could help meet demands changing climate.

Language: Английский

Citations

4