Creative BioMart is a well-known expert specializing in the application of advanced cell-free display technology to accelerate the release of protein engineering. With years of experience, we are able to provide noncovalent DNA display system service based on cell-free technology to precisely meet customer requirements.

Non-covalent DNA display is a new tool for directed evolution of proteins in vitro, which connects genotype (DNA) and phenotype (protein) in a non-covalent manner. In this system, a DNA construct composed of the coding sequence for the protein of interest, the coding sequence for the DNA replication initiator protein RepA, the CIS element, and the R1 plasmid origin of replication (ori) is transcribed and translated in a standard, coupled in vitro reaction. RepA transiently binds to the CIS element, which directs it to bind noncovalently to the nearby ori. This technology can display and select peptides and proteins from super large libraries.

Services

CIS display is a cell-free in vitro display technology based on linear dsDNA template. Creative BioMart provides a complete non-covalent DNA display platform based on CIS display and in vitro compartmentalization. The RepA we provide is a highly reliable cis-acting DNA binding protein, and libraries encoding >10¹² random 18-mer peptides can be constructed and used to isolate peptides that bind specifically to disparate targets. In addition, the use of in vitro compartmentalization DNA display technology uses FACS to screen adhesives.

The CIS display system is an alternative DNA-based method, which does not require any division of library-encoded nucleic acids. This system allows the evolution of peptides, antibody fragments and protein scaffolds, mainly for therapeutic applications.

Fig 1. The principle of the CIS display technology. (Odegrip R, et al., 2004)

Features of Noncovalent DNA Display System

• Unlike phage display technology, CIS display can use bacterial cell lysates in a purely cell-free manner, which overcomes the limitations of other technologies that need to transfer DNA into cells.
• CIS display operation is simple, can quickly generate and screen larger libraries, more than 10¹³ different peptides library size and associated with their own code to shorten the time from library design to hit identification.
• The peptide fused with RepA is fully and effectively connected to its own DNA. In the test using peptide tags, the use of anti-peptide antibodies can recover more than 40% of the DNA template.
• The CIS display system can display different peptide topologies and proteins up to 90 kDa, including antibody fragments and protein scaffolds (such as centyrin scaffolds).

Creative BioMart has been continuously developing cutting-edge cell-free display technology, and is committed to providing a flexible and efficient customized noncovalent DNA display system service. Based on the protein engineering platform, we have successfully constructed an impeccable cell-free surface display library of various types. We are proud to be able to screen and select proteins with the required characteristics from a large number of variants to best accelerate the research of life sciences. If you are interested in our services, please do not hesitate to contact us for more information.

Reference

1. Odegrip R, Coomber D, et al. (2004) CIS display: In vitro selection of peptides from libraries of protein-DNA complexes. Proc Natl Acad Sci U S A. 101(9): 2806-2810.;

• In Vitro Compartmentalization Displays Service
• Covalent DNA Displays Service
• mRNA Displays Service
• Ribosome Displays Service