title: A multicolor panel of TALE-KRAB based transcriptional repressor vectors enabling knockdown of multiple gene targets.
abstract: Stable and efficient knockdown of multiple gene targets is highly desirable for dissection of molecular pathways. Because it allows sequence-specific DNA binding, transcription activator-like effector (TALE) offers a new genetic perturbation technique that allows for gene-specific repression. Here, we constructed a multicolor lentiviral TALE-Kruppel-associated box (KRAB) expression vector platform that enables knockdown of multiple gene targets. This platform is fully compatible with the Golden Gate TALEN and TAL Effector Kit 2.0, a widely used and efficient method for TALE assembly. We showed that this multicolor TALE-KRAB vector system when combined together with bone marrow transplantation could quickly knock down c-kit and PU.1 genes in hematopoietic stem and progenitor cells of recipient mice. Furthermore, our data demonstrated that this platform simultaneously knocked down both c-Kit and PU.1 genes in the same primary cell populations. Together, our results suggest that this multicolor TALE-KRAB vector platform is a promising and versatile tool for knockdown of multiple gene targets and could greatly facilitate dissection of molecular pathways.
authors.affiliation: Division of Hematology/Oncology, Department of Medicine, University of Illinois College of Medicine at Chicago, Chicago, IL 60612, USA.
authors.forename: Zhonghui
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: Winsor School, Boston, MA 02215, USA.
authors.forename: Elise
authors.initials: E
authors.lastname: Wu
authors.affiliation: Division of Hematology/Oncology, Department of Medicine, University of Illinois College of Medicine at Chicago, Chicago, IL 60612, USA.
authors.forename: Zhijian
authors.initials: Z
authors.lastname: Qian
authors.affiliation: Division of Hematology/Oncology, Department of Medicine, University of Illinois College of Medicine at Chicago, Chicago, IL 60612, USA.
authors.forename: Wen-Shu
authors.initials: WS
authors.lastname: Wu
mesh_terms: D003116:Color; D055785:Gene Knockdown Techniques / Q000379:methods*; D018390:Gene Targeting / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics*; D057809:HEK293 Cells; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D012097:Repressor Proteins / Q000235:genetics*; D014158:Transcription, Genetic / Q000235:genetics*
chemical_list: D012097:Repressor Proteins