A CRISPR/Cas9 system adapted for gene editing in marine algae

Area of Science:

• Molecular Biology
• Marine Biology
• Biotechnology

Background:

• Microalgae are crucial for biotechnology and ecological research.
• Targeted gene editing in microalgae remains challenging.
• Developing efficient genetic tools for microalgae is essential.

Purpose of the Study:

• To demonstrate the efficacy of CRISPR/Cas9 for generating targeted gene mutations in microalgae.
• To establish a robust method for screening and validating CRISPR/Cas9-induced mutants.
• To provide a versatile vector system for gene editing in Phaeodactylum tricornutum.

Main Methods:

• Utilized the CRISPR/Cas9 gene editing system.
• Employed a novel vector design for easy construct adaptation.
• Screened for mutants using high-resolution melting (HRM) based PCR assays.
• Confirmed mutations via DNA sequencing and functional analyses.

Main Results:

• Successfully generated stable, targeted gene mutations in the marine diatom Phaeodactylum tricornutum.
• Demonstrated the efficiency and ease of the CRISPR/Cas9 system in microalgae.
• Validated the effectiveness of HRM-PCR for rapid mutant screening.

Conclusions:

• CRISPR/Cas9 is a powerful and efficient tool for genetic engineering in microalgae.
• The developed vector system and screening methods facilitate rapid generation and validation of mutants.
• This technology opens new avenues for functional genomics and synthetic biology in microalgae.