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Appl Microbiol Biotechnol. 2019 Apr;103(8):3239-3248. doi: 10.1007/s00253-019-09726-x. Epub 2019 Mar 15.

Application of the CRISPR/Cas system for genome editing in microalgae.

Author information

1
College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, No. 30 South Puzhu Road, Nanjing, 211816, People's Republic of China.
2
Jiangsu National Synergetic Innovation Center for Advanced Materials (SICAM), No. 30 South Puzhu Road, Nanjing, 211816, People's Republic of China.
3
School of Pharmaceutical Sciences, Nanjing Tech University, No. 30 South Puzhu Road, Nanjing, 211816, People's Republic of China.
4
State Key Laboratory of Materials-Oriented Chemical Engineering, Nanjing Tech University, No. 5 Xinmofan Road, Nanjing, 210009, People's Republic of China.
5
School of Food Science and Pharmaceutical Engineering, Nanjing Normal University, Wenyuan Road, Nanjing, 210023, People's Republic of China.
6
College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, No. 30 South Puzhu Road, Nanjing, 211816, People's Republic of China. renlujing@njtech.edu.cn.
7
Jiangsu National Synergetic Innovation Center for Advanced Materials (SICAM), No. 30 South Puzhu Road, Nanjing, 211816, People's Republic of China. renlujing@njtech.edu.cn.

Abstract

Microalgae are arguably the most abundant single-celled eukaryotes and are widely distributed in oceans and freshwater lakes. Moreover, microalgae are widely used in biotechnology to produce bioenergy and high-value products such as polyunsaturated fatty acids (PUFAs), bioactive peptides, proteins, antioxidants and so on. In general, genetic editing techniques were adapted to increase the production of microalgal metabolites. The main genome editing tools available today include zinc finger nucleases (ZFNs), transcriptional activator-like effector nucleases (TALENs), and the clustered regularly interspaced short palindromic repeats (CRISPR)/Cas nuclease system. Due to its high genome editing efficiency, the CRISPR/Cas system is emerging as the most important genome editing method. In this review, we summarized the available literature on the application of CRISPR/Cas in microalgal genetic engineering, including transformation methods, strategies for the expression of Cas9 and sgRNA, the CRISPR/Cas9-mediated gene knock-in/knock-out strategies, and CRISPR interference expression modification strategies.

KEYWORDS:

CRISPR/Cas; Genome editing; Microalgae; Transformation; sgRNA

PMID:
30877356
DOI:
10.1007/s00253-019-09726-x

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