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ARS Home » Midwest Area » Columbia, Missouri » Plant Genetics Research » Research » Research Project #442519

Research Project: Validate Causative Mutations in Agriculturally-Important Vertebrates

Location: Plant Genetics Research

Project Number: 5070-31320-001-005-S
Project Type: Non-Assistance Cooperative Agreement

Start Date: Aug 15, 2022
End Date: Aug 14, 2025

1) Using gene editing techniques, systematically alter the DNA sequence predicted to cause beneficial changes in pig and poultry production and to test these changes in vitro and, where warranted, in vivo. 2) Test the effects of sequence modifications for on- and off-target effects.

In order to identify targets to further improve livestock animal welfare and production, we have identified genes or QTLs through: 1) genome wide association studies that are associated with positive traits; 2) known genes that are associated with positive traits in other species; and 3) known biological pathways that are associated with positive animal traits. Our goal is to first validate the role the gene plays in the cellular phenotype in vitro before producing gene edited animals. To achieve this goal, both pig and poultry primary cell lines and organoid systems will be established from multiple tissue types. Then, each cell line will be characterized by RNA-sequencing and by using a proteomics biomarker approach. A comparison between our primary cell lines, commercially available cell lines (if available), and cells collected from whole tissue will be completed to understand how well our cell cultures mimic the in vivo tissue. Our second goal is to decrease off-target effects in gene edited cells. This will be accomplished by first establishing the variability in our current transfection system by using a guide that has known off-targeting events.