Identification of ligands that enable endocytosis, using in vivo manipulation of neuronal fibers

摘要

In vivo screening is used to identify and isolate ligands that drive endocytosis (internalisation) of molecules into animal cells. These ligands can transport passenger molecules (drug or diagnostic compounds, genetic vectors, etc.) into targeted classes of cells. A population of candidate ligands, such as a phage display or combinatorial library, is placed in a rat leg, in contact with a sciatic nerve bundle, and a ligature is tightened around the same nerve bundle at the hip. After a delay, to enable ligands that bind to endocytotic receptors on the nerve fibers to be internalised and transported within the fibers, fiber segments are harvested from the ligature site in the hip. Ligands that entered the harvested nerve segments can be isolated, sequenced, reproduced, etc. If desired, rats can be transformed to express human endocytotic receptors, to allow selection of ligands that will be transported into targeted human cells.