Novel Promoters Derived from Chinese Hamster Ovary Cells via In Silico and In Vitro Analysis

Area of Science:

• Biotechnology
• Molecular Biology
• Cell Engineering

Background:

• Industrial recombinant protein production relies on high gene expression, often using viral promoters.
• Viral promoters can cause cellular stress and epigenetic silencing due to uncontrolled gene expression.
• Endogenous promoters offer potential for better cellular state response and reduced stress.

Purpose of the Study:

• To identify and characterize Chinese hamster ovary (CHO) endogenous promoters and enhancers.
• To evaluate their strength and stability for recombinant protein production.
• To explore strategies for enhancing promoter activity.

Main Methods:

• Identification of CHO endogenous promoters and enhancers using histone marks and chromatin states.
• Ranking based on expression levels and testing normalized promoter strength.
• Truncation of promoters and combination with enhancers to optimize activity.

Main Results:

• Novel CHO endogenous promoters and enhancers were identified.
• Promoter activity was enhanced up to threefold through truncation and enhancer combination.
• The strongest CHO promoter demonstrated greater stability than the human cytomegalovirus promoter with enhancer.

Conclusions:

• CHO endogenous promoters are promising alternatives to viral promoters for recombinant protein production.
• These promoters offer reduced cellular stress and improved stability.
• Further research into promoter functionality is needed for advanced cell engineering applications.