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The liver performs a number of vital functions in the chicken. In order to identify unique gene expression patterns and link them to potential functions in the chicken liver, genes enriched in the liver of chickens needed to be investigated in a comparative manner. In this study, 41 liver-enriched genes were identified through chicken microarray, and many of them were validated through comparative analysis of mice and humans. Thirteen of them were unique in chickens, and their liver enhancement was confirmed by reverse transcription PCR. Furthermore, the expression of those 13 chicken liver-enriched genes was investigated, in response to nutritional and physiological challenges. Real-time PCR revealed that expression of PIT54 (P < 0.01), phosphoribosyl pyrophosphate synthetase 2 (PRPS2) (P < 0.05), sulfotransferase (SULT) (P < 0.05), and cytochrome P450 family 2 subfamily C, polypeptide 18 (CYP2C18) (P < 0.05) were significantly decreased in the liver during fasting compared to ad libitum control. During the post-laying stage, expression of GAL8 was significantly increased (P < 0.01), but CYP2C18 expression was significantly reduced (P < 0.05). Liver-enriched genes that were identified in this study and their expression patterns under fasting and the post-laying stage will serve as future targets to gain a better understanding of liver physiology, function and development in poultry.

Genetic selection of quail for a low body weight for more than 80 generations established a low-weight (LW) Japanese quail line that has been previously characterized to have a muscle hypoplasia phenotype. The aim of this study is to investigate the relationship of temporal expression levels of myostatin (Mstn) and myogenic regulatory factors (MRFs) with hypoplastic muscle growth in the LW line. During embryonic day (E) 13 to 15, gain of embryo weight was 2-fold lower (P

Soon after RNA-guided Cas9 (CRISPR-associated protein 9) endonuclease opened a new era of targeted genome editing, the CRISPR/Cas9 platform began to be extensively used to modify genes in various types of cells and organisms. However, successful CRISPR/Cas9-mediated insertion/deletion (indel) mutation remains to be demonstrated in avian cell lines. The objective of this study was to design a poultry-specific CRISPR/Cas9 system to efficiently introduce targeted deletion mutation in chromosomes of the quail muscle clone 7 (QM7) cell line using a customized quail CRISPR vector. In this study, two avian-specific promoters, quail 7SK (q7SK) promoter and CBh promoter, the hybrid form of cytomegalovirus and chicken β-actin promoters, were cloned into a CRISPR vector for the expression of guide RNA and Cas9 protein, respectively. Then, guide RNA, which was designed to target 20-base pair (bp) nucleotides in the quail melanophilin (MLPH) locus, was ligated to the modified CRISPR vector and transfected to QM7 cells. Our results showed multiple indel mutations in the quail MLPH locus in nearly half of the alleles being tested, suggesting the high efficiency of the system for targeted gene modification. The new CRISPR vector developed from this study has the potential application to generate knockout avian cell lines and knockout poultry.