Gateway® Entry Clones

• RNAi Expression - Efficient delivery and long-term stable or transient shRNA expression in any mammalian cell types with Lentiviral and Adenoviral Gateway® vectors
• Viral Expression - High level, stable and transient gene expression in any mammalian cell type
• In Vitro Protein Synthesis - Directly synthesize high yields of recombinant protein in a single reaction tube in just 2 hours without special equipment. Scale the reaction to achieve microgram to milligram levels of protein
• In Vivo Biotinylation - Easy efficient method for expressing, purifying and detecting biotinylated recombinant proteins.
• Tag-On-Demand ™ Technology - Produce native or tagged proteins from one vector.

The three possible methods that lead to the Entry clone are depicted below: A. BP cloning, B. TOPO® cloning and C. restriction enzyme and ligase cloning. Red arrows represent the fragment of interest. Adapted from Katzen, F. 2007) Expert Opin Drug Discov 2(4): 571–589.

pENTR/D-TOPO® vectors take advantage of fast, efficient Directional TOPO® cloning that delivers your insert in the correct orientation for expression. These vectors contain the necessary attL sequences for recombination into any Destination vector and certain versions carry a TEV protease cleavage site for producing native proteins after expression (Figure 1).

Figure 1. Several pENTR™ vectors are available for Directional TOPO® cloning and direct access to the multitude of Gateway® expression vectors.

The pCR®8/GW/TOPO® vector enables efficient TOPO®-TA cloning. These vectors easily facilitate multipurpose use: rapid recombination into a variety of Gateway® Destination vectors, convenient sequencing, robust selection in E. coli with spectinomycin resistance, and easy exicision of insert DNA with flanking EcoR I sites (Figure 2).

Figure 2. The pCR®8/GW/TOPO® Entry vector allows TOPO® TA Cloning® for multiple downstream applications.