Format

Send to

Choose Destination
Sci Rep. 2016 Jul 28;6:30620. doi: 10.1038/srep30620.

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

Author information

1
Department of Life Science, Hanyang University, Seoul, South Korea.
2
Department of Chemistry, Hanyang University, Seoul, South Korea.
3
Department of Chemical and Biological Engineering, Korea University, Seoul, South Korea.
4
Department of Plant and Microbial Biology, University of California, Berkeley, CA 94720-3102, USA.
5
Center for Genome Engineering, Institute for Basic Science, Seoul, South Korea.
6
Department of Chemistry, Seoul National University, Seoul, South Korea.

Abstract

Microalgae are versatile organisms capable of converting CO2, H2O, 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.

PMID:
27466170
PMCID:
PMC4964356
DOI:
10.1038/srep30620
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for Nature Publishing Group Icon for PubMed Central
Loading ...
Support Center