Invitrogen™ Dynabeads™ Streptavidin for In Vitro Transcription

Description

Dynabeads Streptavidin for In Vitro Transcription offer an innovative technology for simple, flexible, and automatable mRNA synthesis, in scales ranging from micrograms to grams. They are particularly suitable where flexibility and high quality are required, such as in the discovery phase of RNA-based vaccines and therapeutics development.

The 1-μm monosized, monodispersed, paramagnetic beads are covalently coupled with streptavidin molecules, for binding to biotinylated DNA template. The workflow is designed to minimize the effort and cost of plasmid preparation, while enabling reuse of template-bead complex in successive solid-phase in vitro transcription (IVT) reactions (figure below).

Features of Dynabeads Streptavidin for In Vitro Transcription include:

• Flexibility – choose DNA template for IVT with flexibility of design and amplification
• Efficiency – reuse DNA template and beads at least six times to help reduce cost and eliminate/minimize plasmid prep hands-on time and risk of bacterial host DNA transfer
• Scalability – swap between manual and automated workflows to match required scale (micrograms to grams of mRNA) and throughput
• Quality – help ensure a safe and flexible supply of magnetic beads to navigate from RNA-based drug discovery research to commercial manufacturing with counterpart solutions (available via the OEM supply) without incurring extra time or costs from switching technology or raw material

Overview of the workflow
A biotinylated template is made by PCR amplification of the target sequence in a plasmid construct or in a synthetic DNA construct using a biotinylated forward primer and a non-biotinylated reverse primer. As an alternative to PCR amplification, a linearized plasmid can be biotinylated by a fill-in reaction of biotin-dUTP in the 5' overhang sequence, provided the correct plasmid design. The biotinylated template is then immobilized directly to the Dynabeads Streptavidin for In Vitro Transcription beads without the need for prior purification. The bead-bound template is used directly in the IVT, then removed from the synthesized mRNA by magnetic separation (Figure 1). The template can be reused (at least six times) in consecutive IVT reactions (figure below).

Solid-phase IVT is compatible with commonly used reagents such as MEGAscript T7 Transcription Kit, mMESSAGE mMACHINE T7 Transcription Kit, and mMESSAGE mMACHINE T7 ULTRA Transcription Kit, based on your needs and strategies for capping and polyadenylation.

The beads are offered here as a stand-alone product. They are also included in the Dynabeads In Vitro Transcription and RNA Purification Kit.

Automation-ready for use with KingFisher purification systems
The 1-μm magnetic beads are ideal for high-throughput screening. The process can be automated using liquid handlers or any of our KingFisher sample purification systems. The scripts for KingFisher Flex and KingFisher Apex automated purification workflows are available from our website.

Commercial supply
To learn more about counterpart products and support documentation available for GMP-compliant manufacturing of mRNA vaccines and therapeutics, visit: thermofisher.com/dynabeadsmrna