PRK introns provide modular tools for enhanced transgene expression in Chlamydomonas reinhardtii
Chlamydomonas reinhardtii is an attractive chassis for photosynthetic biotechnology, but reliable nuclear transgene expression remains a major limitation. Despite advances in genetic engineering, achieving robust nuclear transgene expression in this organism remains challenging. One effective strategy is the introduction of introns into transgene sequences to trigger Intron-Mediated Enhancement…
The study explores how introns from the PRK gene can be used to enhance transgene expression in the single-celled organism Chlamydomonas reinhardtii. This organism is a popular choice for photosynthetic biotechnology, but getting genes to express properly inside its nucleus is still a major hurdle. Previous work showed that genes with endogenous introns often performed better than those without, such as the phosphoribulokinase (PRK) gene.
Researchers specifically examined the first intron of the RBCS2 gene (RBCS2i1) as a benchmark for IME activity, the common term for this enhanced expression. Comparing PRKi1 to the original RBCS2i1 intron, the studies found that PRKi1 could boost expression by up to 35 times. These findings were tested using different reporting systems, genetic backgrounds, and promoter-terminator setups, consistently revealing significant expression improvements.
Crucially, PRKi1 was shown to be effective even for a transgene like patchoulol synthase, a complex one that generally needs introns to be expressed at all. This makes PRKi1 and its close relative PRKi2 look like promising, adaptable tools for boosting gene expression in C. reinhardtii for a variety of synthetic biology and biotechnological applications.
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