ArticleMicrobial biotechnology2024
High-efficiency genome editing by Cas12a ribonucleoprotein complex in Euglena gracilis.
Article in Microbial biotechnology, 2024. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 10 papers.
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10 citing papers in PubMed.
- Mapping the proteome landscape inPlant signaling & behavior · 2026Article
- Systematic Profiling of ssODN Determinants Establishes Optimized LbCas12a-Mediated HDR for Precise Genome Editing inJournal of fungi (Basel, Switzerland) · 2026Article
- Affordable CRISPR RNP-Based Genome Editing in Euglena gracilis.Current protocols · 2026Article
- CRISPR-enhanced microalgae biosynthesis: a promising approach for future functional feed ingredients.Journal of animal science and biotechnology · 2026Review
- Acceleration and Light-Induced Changes in Cytosolic cAMP Concentration inBiomolecules · 2026Article
- Engineering Wax Ester Composition in Euglena gracilis Using Genome Editing.Methods in molecular biology (Clifton, N.J.) · 2026Article
- Harnessing microalgae for bioproducts: innovations in synthetic biology.World journal of microbiology & biotechnology · 2025Review
- Progress and challenges in CRISPR/Cas applications in microalgae.Journal of microbiology (Seoul, Korea) · 2025Review
- Changes in Gravitaxis and Gene-Expression in anBiomolecules · 2024Article
- High-efficiency genome editing by Cas12a ribonucleoprotein complex in Euglena gracilis.Microbial biotechnology · 2024Article
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5 authors.
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Abstract
Transgene-free genome editing based on clustered regularly interspaced short palindromic repeats (CRISPR) technology is key to achieving genetic engineering in microalgae for basic research and industrial applications. Euglena gracilis, a unicellular phytoflagellate microalga, is a promising biomaterial for foods, feeds, cosmetics and biofuels. However, methods for the genetic manipulation of E. gracilis are still limited. Here, we developed a high-efficiency, transgene-free genome editing method for E. gracilis using Lachnospiraceae bacterium CRISPR-associated protein 12a (LbCas12a) ribonucleoprotein (RNP) complex, which complements the previously established Cas9 RNP-based method. Through the direct delivery of LbCas12a-containing RNPs, our method reached mutagenesis rates of approximately 77.2-94.5% at two different E. gracilis target genes, Glucan synthase-like 2 (EgGSL2) and a phytoene synthase gene (EgcrtB). Moreover, in addition to targeted mutagenesis, we demonstrated efficient knock-in and base editing at the target site using LbCas12a-based RNPs with a single-stranded DNA donor template in E. gracilis. This study extends the genetic engineering capabilities of Euglena to accelerate its basic use for research and engineering for bioproduction.
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