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

Kwangryul Baek, Duk Hyoung Kim, Jooyeon Jeong, Sang Jun Sim, Anastasios Melis, Jin Soo Kim, Eonseon Jin, Sangsu Bae

Research output: Contribution to journalArticlepeer-review

239 Citations (Scopus)


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.

Original languageEnglish
Article number30620
JournalScientific reports
Publication statusPublished - 2016 Jul 28

Bibliographical note

Funding Information:
This work was supported by grants from Institute for Basic Science (IBS-R021-D1) to J.-S.K.The Korea CCS R&D Center (KCRC) (NRF-2014M1A8A1049273) to E.J. and the Plant Molecular Breeding Center of Next Generation BioGreen 21 Program (PJ01119201) to S.B.

ASJC Scopus subject areas

  • General


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