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Main Authors: Murata, Satoshi, Kushiyama, Natsuno, Yabu, Yusuke, Watanabe, Kahori, Fujii, Taiga, Yasui, Rein, Nojima, Daisuke, Maeda, Yoshiaki, Yoshino, Tomoko, Matsuda, Yusuke, Tanaka, Tsuyoshi
Format: Artículo científico
Language:en
Published: Journal of bioscience and bioengineering 2025
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Online Access:https://pubmed.ncbi.nlm.nih.gov/40803956/
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author Murata, Satoshi
Kushiyama, Natsuno
Yabu, Yusuke
Watanabe, Kahori
Fujii, Taiga
Yasui, Rein
Nojima, Daisuke
Maeda, Yoshiaki
Yoshino, Tomoko
Matsuda, Yusuke
Tanaka, Tsuyoshi
author_facet Murata, Satoshi
Kushiyama, Natsuno
Yabu, Yusuke
Watanabe, Kahori
Fujii, Taiga
Yasui, Rein
Nojima, Daisuke
Maeda, Yoshiaki
Yoshino, Tomoko
Matsuda, Yusuke
Tanaka, Tsuyoshi
Murata, Satoshi
Kushiyama, Natsuno
Yabu, Yusuke
Watanabe, Kahori
Fujii, Taiga
Yasui, Rein
Nojima, Daisuke
Maeda, Yoshiaki
Yoshino, Tomoko
Matsuda, Yusuke
Tanaka, Tsuyoshi
collection PubMed - marine biology
contents Establishment of genome editing techniques in the marine oleaginous diatom Fistulifera solaris for improved oil accumulation. Murata, Satoshi Kushiyama, Natsuno Yabu, Yusuke Watanabe, Kahori Fujii, Taiga Yasui, Rein Nojima, Daisuke Maeda, Yoshiaki Yoshino, Tomoko Matsuda, Yusuke Tanaka, Tsuyoshi Diatoms Gene Editing CRISPR-Cas Systems Biofuels Lipase Gene Knockout Techniques Oils Microalgae Triglycerides Lipid Metabolism Biofuel production using microalgae has attracted considerable attention owing to high growth rate and lipid accumulation properties. However, further enhancement in lipid productivity is required to render this economically feasible. CRISPR/Cas9, which is one of the powerful genome editing tools, is an essential technique that may solve this problem. The marine diatom Fistulifera solaris JPCC DA0580 is a promising candidate of the biofuel production, since it accumulates significant amount of lipids. However, genome editing techniques have not yet been established for F. solaris, which prevent the construction of valuable strains. In this study, CRISPR/Cas9-mediated specific gene knockout technique was established in F. solaris, through targeting adenine phosphoribosyl transferase gene (apt) and triacylglycerol (TAG) lipase gene (tgl1). Mutations in the target sequence were detected in apt- and tgl1-edited mutants. Moreover, the mutants showed distinct phenotypes, such as suppression of TAG degradation and resistance to 2-fluoroadenine. These results indicate the successful demonstration of CRISPR/Cas9-mediated genome editing in the oleaginous marine diatom F. solaris. Furthermore, oil degradation was successfully suppressed by knocking-out tgl1. The CRISPR/Cas9-mediated genome editing established in this study provides key molecular tools for both the basic biology and the future biotechnological applications of F. solaris, such as biofuel production.
format Artículo científico
id pubmed_40803956
institution PubMed
language en
publishDate 2025
publisher Journal of bioscience and bioengineering
record_format pubmed
spellingShingle Establishment of genome editing techniques in the marine oleaginous diatom Fistulifera solaris for improved oil accumulation.
Murata, Satoshi
Kushiyama, Natsuno
Yabu, Yusuke
Watanabe, Kahori
Fujii, Taiga
Yasui, Rein
Nojima, Daisuke
Maeda, Yoshiaki
Yoshino, Tomoko
Matsuda, Yusuke
Tanaka, Tsuyoshi
Diatoms
Gene Editing
CRISPR-Cas Systems
Biofuels
Lipase
Gene Knockout Techniques
Oils
Microalgae
Triglycerides
Lipid Metabolism
Establishment of genome editing techniques in the marine oleaginous diatom Fistulifera solaris for improved oil accumulation. Murata, Satoshi Kushiyama, Natsuno Yabu, Yusuke Watanabe, Kahori Fujii, Taiga Yasui, Rein Nojima, Daisuke Maeda, Yoshiaki Yoshino, Tomoko Matsuda, Yusuke Tanaka, Tsuyoshi Diatoms Gene Editing CRISPR-Cas Systems Biofuels Lipase Gene Knockout Techniques Oils Microalgae Triglycerides Lipid Metabolism Biofuel production using microalgae has attracted considerable attention owing to high growth rate and lipid accumulation properties. However, further enhancement in lipid productivity is required to render this economically feasible. CRISPR/Cas9, which is one of the powerful genome editing tools, is an essential technique that may solve this problem. The marine diatom Fistulifera solaris JPCC DA0580 is a promising candidate of the biofuel production, since it accumulates significant amount of lipids. However, genome editing techniques have not yet been established for F. solaris, which prevent the construction of valuable strains. In this study, CRISPR/Cas9-mediated specific gene knockout technique was established in F. solaris, through targeting adenine phosphoribosyl transferase gene (apt) and triacylglycerol (TAG) lipase gene (tgl1). Mutations in the target sequence were detected in apt- and tgl1-edited mutants. Moreover, the mutants showed distinct phenotypes, such as suppression of TAG degradation and resistance to 2-fluoroadenine. These results indicate the successful demonstration of CRISPR/Cas9-mediated genome editing in the oleaginous marine diatom F. solaris. Furthermore, oil degradation was successfully suppressed by knocking-out tgl1. The CRISPR/Cas9-mediated genome editing established in this study provides key molecular tools for both the basic biology and the future biotechnological applications of F. solaris, such as biofuel production.
title Establishment of genome editing techniques in the marine oleaginous diatom Fistulifera solaris for improved oil accumulation.
topic Diatoms
Gene Editing
CRISPR-Cas Systems
Biofuels
Lipase
Gene Knockout Techniques
Oils
Microalgae
Triglycerides
Lipid Metabolism
url https://pubmed.ncbi.nlm.nih.gov/40803956/