title: Transcription activator-like effector hybrids for conditional control and rewiring of chromosomal transgene expression.
abstract: The ability to conditionally rewire pathways in human cells holds great therapeutic potential. Transcription activator-like effectors (TALEs) are a class of naturally occurring specific DNA binding proteins that can be used to introduce targeted genome modifications or control gene expression. Here we present TALE hybrids engineered to respond to endogenous signals and capable of controlling transgenes by applying a predetermined and tunable action at the single-cell level. Specifically, we first demonstrate that combinations of TALEs can be used to modulate the expression of stably integrated genes in kidney cells. We then introduce a general purpose two-hybrid approach that can be customized to regulate the function of any TALE either using effector molecules or a heterodimerization reaction. Finally, we demonstrate the successful interface of TALEs to specific endogenous signals, namely hypoxia signaling and microRNAs, essentially closing the loop between cellular information and chromosomal transgene expression.
authors.affiliation: Bioengineering Department, The University of Texas at Dallas , 800 West Campbell Road, Richardson, TX 75080, USA.
authors.forename: Yi
authors.initials: Y
authors.lastname: Li
authors.forename: Richard
authors.initials: R
authors.lastname: Moore
authors.forename: Michael
authors.initials: M
authors.lastname: Guinn
authors.forename: Leonidas
authors.initials: L
authors.lastname: Bleris
mesh_terms: D002460:Cell Line / Q000187:drug effects; D002678:Chimera; D004318:Doxycycline / Q000494:pharmacology; D005786:Gene Expression Regulation; D005818:Genetic Engineering / Q000379:methods*; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000235:genetics; D035683:MicroRNAs / Q000378:metabolism; D011401:Promoter Regions, Genetic; D015202:Protein Engineering / Q000379:methods; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000378:metabolism; D020218:Response Elements; D019076:Transgenes
chemical_list: C497442:HIF1A protein, human; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; C505362:MIRN16 microRNA, human; C524941:MIRN17 microRNA, human; D035683:MicroRNAs; D011993:Recombinant Fusion Proteins; D004318:Doxycycline