Efficiently Editing Multiple Duplicated Homeologs and Alleles for Recurrent Polyploids

Area of Science:

• Genomics
• Evolutionary Biology
• Molecular Biology

Background:

• Studying the evolutionary fate of duplicated genes in polyploids is challenging.
• Distinguishing homeologs and alleles in duplicated gene regions is complex.

Purpose of the Study:

• To detail methods for identifying and analyzing homeologs and alleles of duplicated genes.
• To investigate functional divergence using gene editing in recurrent polyploids.
• To demonstrate the utility of CRISPR/Cas9 for polyploid genome research.

Main Methods:

• Utilized CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats/CRISPR-associated system 9) gene editing.
• Developed procedures for identifying and characterizing homeologs and alleles.
• Constructed multiple knockout mutant lines in a recurrent polyploid fish model.

Main Results:

• Successfully identified and analyzed homeologs and alleles of duplicated genes.
• Demonstrated functional divergence through gene editing.
• Created knockout mutants targeting single or multiple homeologs/alleles efficiently.

Conclusions:

• The described gene editing approach is effective for targeting multiple homeologs and alleles in recurrent polyploid genomes.
• This methodology facilitates research into the evolutionary dynamics of duplicated genes in polyploids.