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

被引：221

作者：

Baek, Kwangryul ^{[1
]}

Kim, Duk Hyoung ^{[2
]}

Jeong, Jooyeon ^{[1
]}

Sim, Sang Jun ^{[3
]}

Melis, Anastasios ^{[4
]}

Kim, Jin-Soo ^{[5
,6
]}

Jin, EonSeon ^{[1
]}

Bae, Sangsu ^{[2
]}

机构：

[1] Hanyang Univ, Dept Life Sci, Seoul, South Korea

[2] Hanyang Univ, Dept Chem, Seoul, South Korea

[3] Korea Univ, Dept Chem & Biol Engn, Seoul, South Korea

[4] Univ Calif Berkeley, Dept Plant & Microbial Biol, Berkeley, CA 94720 USA

[5] Inst for Basic Sci Korea, Ctr Genome Engn, Seoul, South Korea

[6] Seoul Natl Univ, Dept Chem, Seoul, South Korea

来源：

SCIENTIFIC REPORTS | 2016年 / 6卷

基金：

新加坡国家研究基金会;

关键词：

CAS9; EXPRESSION; ZEAXANTHIN; DELIVERY; GENES; GUIDE;

D O I：

10.1038/srep30620

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

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.

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页数：7

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