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 journalArticle

88 Citations (Scopus)

Abstract

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
Volume6
DOIs
Publication statusPublished - 2016 Jul 28

Fingerprint

Clustered Regularly Interspaced Short Palindromic Repeats
Microalgae
Chlamydomonas reinhardtii
Gene Knockout Techniques
Ribonucleoproteins
DNA
Guide RNA
Sunlight
Carbon Monoxide
Mutagenesis
Biomass
Plasmids

ASJC Scopus subject areas

  • General

Cite this

DNA-free two-gene knockout in Chlamydomonas reinhardtii via CRISPR-Cas9 ribonucleoproteins. / Baek, Kwangryul; Kim, Duk Hyoung; Jeong, Jooyeon; Sim, Sang Jun; Melis, Anastasios; Kim, Jin Soo; Jin, Eonseon; Bae, Sangsu.

In: Scientific Reports, Vol. 6, 30620, 28.07.2016.

Research output: Contribution to journalArticle

Baek, Kwangryul ; Kim, Duk Hyoung ; Jeong, Jooyeon ; Sim, Sang Jun ; Melis, Anastasios ; Kim, Jin Soo ; Jin, Eonseon ; Bae, Sangsu. / DNA-free two-gene knockout in Chlamydomonas reinhardtii via CRISPR-Cas9 ribonucleoproteins. In: Scientific Reports. 2016 ; Vol. 6.
@article{cdce0515f3154dd2a486ae1bfafff8da,
title = "DNA-free two-gene knockout in Chlamydomonas reinhardtii via CRISPR-Cas9 ribonucleoproteins",
abstract = "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.",
author = "Kwangryul Baek and Kim, {Duk Hyoung} and Jooyeon Jeong and Sim, {Sang Jun} and Anastasios Melis and Kim, {Jin Soo} and Eonseon Jin and Sangsu Bae",
year = "2016",
month = "7",
day = "28",
doi = "10.1038/srep30620",
language = "English",
volume = "6",
journal = "Scientific Reports",
issn = "2045-2322",
publisher = "Nature Publishing Group",

}

TY - JOUR

T1 - DNA-free two-gene knockout in Chlamydomonas reinhardtii via CRISPR-Cas9 ribonucleoproteins

AU - Baek, Kwangryul

AU - Kim, Duk Hyoung

AU - Jeong, Jooyeon

AU - Sim, Sang Jun

AU - Melis, Anastasios

AU - Kim, Jin Soo

AU - Jin, Eonseon

AU - Bae, Sangsu

PY - 2016/7/28

Y1 - 2016/7/28

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=84979939297&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84979939297&partnerID=8YFLogxK

U2 - 10.1038/srep30620

DO - 10.1038/srep30620

M3 - Article

C2 - 27466170

AN - SCOPUS:84979939297

VL - 6

JO - Scientific Reports

JF - Scientific Reports

SN - 2045-2322

M1 - 30620

ER -