Engineered cellular pathways for programmed autoregulation of differentiation

摘要

The present invention provides compositions and methods for programming mammalian cells to perform desired functions. In particular, the present invention provides compositions and methods for programming stem cells to differentiate into a desired cell type. A quorum sensing systems that regulates the expression of cell fate regulators is introduced into mammalian host cells, such as stem cells. The quorum sensing systems generally comprises vectors that express the components of a bacterial quorum sensing pathway, including proteins which catalyze the synthesis of an autoinducer and a gene encoding a regulatory partner of the autoinducer, and vectors in which genes encoding cell fate regulators are operably linked to a promoter induced by the autoinducer/regulatory partner complex. The system can also comprise vectors in which genes encoding additional cell fate regulators are operably linked to a promoter that is induced by a factor synthesized in response to a first stage of differentiation, so that a second stage of differentiation is triggered.

主权项

1. A composition comprising one or more recombinant mammalian vectors, wherein the one or more vectors comprise:
a) a first nucleic acid sequence comprising a first promoter that is operably linked to a first DNA sequence encoding a first cell-cell communication molecule, b) a second nucleic acid sequence comprising a second promoter that is operably linked to second DNA sequence encoding a second cell-cell communication molecule that binds with said first cell-cell communication molecule to produce a first binding molecule, c) a third nucleic acid sequence comprising a third promoter that is operably linked to a third DNA sequence encoding a first cell fate regulator protein that induces differentiation of a first cell type into a second cell type that expresses a marker protein of said second cell type, d) a fourth nucleic acid sequence comprising a fourth promoter that is operably linked to a fourth DNA sequence encoding a first repressor, and e) a fifth nucleic acid sequence comprising a fifth promoter that is operably linked to a fifth DNA sequence encoding a first activator, wherein
i) said first repressor represses expression of said first activator, andii) said first activator activates expression of;
1) said first repressor, and2) said first activator.