CRISPR technology is the most robust genome manipulation tool that allows researchers to generate animals with new features. Using CRISPR technology various animals with an improved immune system, disease resistance, and the ability in producing drugs have been generated. In transgenic chicken, under the control of the ovalbumin gene promoter, biopharmaceutical proteins are produced in the oviduct and secreted into the egg white. In this study to exploit the promoter of the ovalbumin gene, we inserted a promoterless reporter gene in the 3’-UTR region of this gene in chicken DF1 cells using CRISPR-HDR. Then, we applied our recently developed CRISPR/dCas9-mediated transposition technology to incorporate a ubiquitous promoter (CMV), upstream of ovalbumin exon 1 and to activate this promoter in DF1 cells. Our results showed that the 3’UTR region of the ovalbumin gene is a suitable candidate to support the expression of the transgene in chicken bioreactors. Also, our introduced dCas9-piggyBac transposition system provided a useful tool for studying transgene expression in non-oviduct cells. These methods provide useful tools for the generation of transgenic chicken bioreactors.

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