DNA-free two-gene knockout in Chlamydomonas reinhardtii via CRISPR-Cas9 ribonucleoproteins

Microalgae are versatile organisms capable of converting CO 2 , H 2 O and sunlight into fuel and chemicals for domestic and industrial consumption. Thus, genetic modifications of microalgae for enhancing photosynthetic productivity and biomass and bio-products generation are crucial for both academi...

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Bibliographic Details
Published in:Scientific reports 2016-07, Vol.6 (1), p.30620, Article 30620
Main Authors: Baek, Kwangryul, Kim, Duk Hyoung, Jeong, Jooyeon, Sim, Sang Jun, Melis, Anastasios, Kim, Jin-Soo, Jin, EonSeon, Bae, Sangsu
Format: Article
Language:English
Subjects:
42/41
45/23
45/41
631/1647/1511
631/449/2491/2046
Algal Proteins - genetics
Chlamydomonas reinhardtii - genetics
CRISPR-Cas Systems
Gene Knockout Techniques - methods
Humanities and Social Sciences
Metabolic Engineering
multidisciplinary
Photosynthesis
Ribonucleoproteins - metabolism
Science
Zeaxanthins - metabolism
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Summary:Microalgae are versatile organisms capable of converting CO 2 , H 2 O and sunlight into fuel and chemicals for domestic and industrial consumption. Thus, genetic modifications of microalgae for enhancing photosynthetic productivity and biomass and bio-products generation are crucial for both academic and industrial applications. However, targeted mutagenesis in microalgae with CRISPR-Cas9 is limited. Here we report, a one-step transformation of Chlamydomonas reinhardtii by the DNA-free CRISPR-Cas9 method rather than plasmids that encode Cas9 and guide RNAs. Outcome was the sequential CpFTSY and ZEP two-gene knockout and the generation of a strain constitutively producing zeaxanthin and showing improved photosynthetic productivity.
ISSN:2045-2322
2045-2322
DOI:10.1038/srep30620