Targeted genome editing in a quail cell line using a customized CRISPR/Cas9 system

Area of Science:

• Genetics
• Molecular Biology
• Biotechnology

Background:

• CRISPR-associated protein 9 (Cas9) enables targeted genome editing.
• CRISPR/Cas9 technology has been widely applied across various organisms.
• Efficient CRISPR/Cas9-mediated gene editing in avian cell lines remains a challenge.

Purpose of the Study:

• To design and validate a poultry-specific CRISPR/Cas9 system.
• To achieve efficient targeted deletion mutations in avian cell lines.
• To develop a customized quail CRISPR vector for gene modification.

Main Methods:

• Utilized avian-specific promoters (quail 7SK and CBh) for gene expression.
• Designed guide RNA targeting the quail melanophilin (MLPH) locus.
• Transfected the modified CRISPR vector into quail muscle clone 7 (QM7) cells.

Main Results:

• Demonstrated high efficiency of targeted gene modification in QM7 cells.
• Observed multiple insertion/deletion (indel) mutations in the quail MLPH locus.
• Nearly half of the tested alleles showed mutations, indicating system efficacy.

Conclusions:

• The developed CRISPR/Cas9 system is highly efficient for targeted gene modification in avian cells.
• The customized quail CRISPR vector shows potential for generating knockout avian cell lines.
• This technology can be applied to create knockout poultry for research and development.