Introns mediate post-transcriptional enhancement of nuclear gene expression in the green microalga Chlamydomonas reinhardtii

PLoS Genet. 2020 Jul 30;16(7):e1008944. doi: 10.1371/journal.pgen.1008944. eCollection 2020 Jul.

Abstract

Efficient nuclear transgene expression in the green microalga Chlamydomonas reinhardtii is generally hindered by low transcription rates. Introns can increase transcript abundance by a process called Intron-Mediated Enhancement (IME) in this alga and has been broadly observed in other eukaryotes. However, the mechanisms of IME in microalgae are poorly understood. Here, we identified 33 native introns from highly expressed genes in C. reinhardtii selected from transcriptome studies as well as 13 non-native introns. We investigated their IME capacities and probed the mechanism of action by modification of splice sites, internal sequence motifs, and position within transgenes. Several introns were found to elicit strong IME and found to be broadly applicable in different expression constructs. We determined that IME in C. reinhardtii exclusively occurs from introns within transcribed ORFs regardless of the promoter and is not induced by traditional enhancers of transcription. Our results elucidate some mechanistic details of IME in C. reinhardtii, which are similar to those observed in higher plants yet underly distinctly different induction processes. Our findings narrow the focus of targets responsible for algal IME and provides evidence that introns are underestimated regulators of C. reinhardtii nuclear gene expression.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chlamydomonas reinhardtii / genetics*
  • Gene Expression Regulation, Plant / genetics
  • Introns / genetics*
  • Microalgae / genetics
  • Promoter Regions, Genetic
  • Protein Processing, Post-Translational / genetics*
  • RNA Splicing / genetics*
  • Transcriptome / genetics

Grants and funding

The authors would like to acknowledge financial support by the European Union’s ERA-NET Cofund on Biotechnologies and the Federal Ministry of Education and Research of Germany (BMBF no. 031B0613) in the framework “MERIT” (to T.B., A.E. and O.K.). The authors would like to acknowledge support for the Article Processing Charge by the Deutsche Forschungsgemeinschaft and the Open Access Publication Fund of Bielefeld University. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.