The poultry industry provides cost-effective, healthy, and protein-enriched food for the growing population and achieving the nutritional security to the country. Excessive abdominal and subcutaneous fat deposition is one of the major setbacks to the poultry industry that reduces carcass yield and feed efficiency. In chicken abdominal fat constitutes 20% of total body fat which make up 2–3% of live weight of the bird. In fatty acid metabolism, acetyl-CoA Carboxylase (ACC) is one of the key enzymes with two isoforms i.e. ACACA and ACACB each of which plays a different role. In chicken, ACACB is involved in the β-oxidation of fatty acids and thereby potentially regulating the quality of meat and egg. The RNAi strategy is widely used for silencing the target gene expression. In this study, we designed five shRNA constructs and identified the most efficient shRNA molecule for silencing the ACACB gene under in vitro chicken embryo myoblast (CEM) primary cell culture system. After knocking down the ACACB gene, for understanding how fatty acid metabolism is regulated, we tracked the expression of key fatty acid metabolism genes like ACACA, FASN, SCD, ELOVL2, and CPT1. Also, checked the expression of immune response genes like IFNA, IFNB, and BLB1 in control as well as ACACB knockdown myoblast cells and observed no significant difference. We observed the down-regulation of key fatty acid metabolism genes along with ACACB, which may leads to the less fat accumulation in CEM cells. We also estimated the cholesterol and triglycerides in control and ACACB knockdown myoblast cells and found a significant difference between control and the knockdown cells. In vitro knockdown of the ACACB gene in a cell culture system by a short hairpin RNA (shRNA) expressing construct would help to produce a knockdown chicken with reduced fat deposition.
