Nils Hansen provides an in-depth overview of the identification of molecular glues using DNA-encoded libraries (DELs) in cells. The presentation begins with an introduction to Vipergen, a company based in Copenhagen that specialises in synthesising and screening DNA-encoded libraries for partners. The company has strategic partnerships in Lithuania and India to expand its library capabilities.
The document highlights the advantages of screening inside cells, which include performing screenings under physiological conditions and eliminating the need for active target proteins. This approach broadens the target base and accelerates the screening process. The fundamental challenge of getting the DNA-encoded library into the cell is addressed by injecting the library into large cells, specifically oocytes from Xenopus Laevis.
The screening process involves generating cDNA of the protein of interest, fusing it to Prey, and injecting it into the cell. After protein expression, the DNA-encoded libraries are injected along with bait DNA. The binding events are then measured by ligating the two pieces of DNA inside the droplet, amplifying the DNA by PCR, and decoding and counting the sequences.
The presentation also discusses the identification of molecular glues in a multiplex fashion by expressing the protein of interest together with E3 ligase. The screening process looks for shared hits in both channels, indicating the presence of molecular glues. The system's effectiveness is demonstrated through various experiments, including the expression of human E3 ligases in frog cells, which are shown to be active.
In summary, Hansen outlines a sophisticated method for identifying molecular glues using DELs in cells. The approach leverages the advantages of physiological screening conditions and multiplexing to identify potential molecular glues efficiently.
