Efficient genome editing in a Mozambique tilapia cell line using CAS ribonucleoprotein complexes

Area of Science:

• Aquaculture and Aquatic Genomics
• Molecular Biology and Gene Editing Technologies

Background:

• Global tilapia aquaculture is significantly impacted by Tilapia lake virus (TiLV).
• The Mozambique tilapia (Oreochromis mossambicus) brain (OmB) cell line is susceptible to TiLV, serving as a valuable in vitro model.
• Efficient genome editing protocols are necessary to utilize OmB cells for genetic improvement in tilapia.

Purpose of the Study:

• To optimize a CRISPR/Cas9 genome editing system for the OmB tilapia cell line.
• To establish a reliable platform for functional genomic studies in tilapia.
• To support the development of tilapia strains resistant to TiLV.

Main Methods:

• Utilized CRISPR/Cas9 ribonucleoprotein (RNP) complexes for genome editing.
• Optimized the genome editing workflow specifically for OmB cells.
• Assessed editing efficiency on two endogenous genes.

Main Results:

• Successfully optimized a CRISPR/Cas9 RNP-based genome editing protocol for OmB cells.
• Achieved high gene editing efficiencies, ranging from 67% to 70% for two targeted genes.
• Established a robust and efficient gene editing workflow for tilapia cell lines.

Conclusions:

• An efficient CRISPR/Cas9 RNP gene editing workflow for OmB tilapia cells has been established.
• This optimized platform will accelerate functional genomic research in tilapia.
• The developed method is expected to aid in breeding TiLV-resistant tilapia for improved aquaculture.