Liver Expression of IGF2 and Related Proteins in ZBED6 Gene-Edited Pig by RNA-Seq

SIMPLE SUMMARY: Zinc finger BED-type containing 6 (ZBED6), as a regulatory factor, has different regulatory mechanisms in animal development. The intron of insulin-like growth factor 2 (IGF2) regulates the development of animal muscle and adipose by combining with the binding site of ZBED6. As a member of the insulin-like growth factor family, IGF2 plays an important role in embryonic growth and development, cell proliferation, muscle growth and genome imprinting. In order to further study the regulatory mechanism of ZBED6 on IGF2, we detected the expression of IGF2 and related genes in ZBED6 single allele knockout (ZBED6-SKO) pig tissues and analyzed differently expressed genes of the transcriptome of ZBED6-SKO pig liver. The results showed that the partial knockout of ZBED6 could affect the secretion of IGF2 in pig liver but had no significant difference at the protein level. This research provides a new idea for the interaction between IGF2 and ZBED6. ABSTRACT: Zinc finger BED-type containing 6 (ZBED6), a highly conservative transcription factor of placental mammals, has conservative interaction of insulin-like growth factor 2 (IGF2) based on the 16 bp binding sites of ZBED6 on the IGF2 sequence. IGF2 is related to embryo growth and cell proliferation. At the same time, its functions in muscle and adipose in mammals have been widely mentioned in recent studies. To further investigate the mechanism of ZBED6 on IGF2, we detected the expression of IGF2 and related genes in ZBED6 single allele knockout (ZBED6-SKO) pig tissues and analyzed the transcriptome of ZBED6-SKO pig liver. Through RNA-seq, we captured nine up-regulated genes and eight down-regulated genes which related to lipid metabolism. The results showed that the mRNA of IGF2 had an upward trend after the partial knockout of ZBED6 in liver and had no significant difference in protein expression of IGF2. In summary, ZBED6-SKO could affect the secretion of IGF2 in pig liver and its own lipid metabolism. Our research has provided basic information for revealing the regulatory mechanism of the interaction between ZBED6 and IGF2 in mammals.