An RNAi-based system for loss-of-function analysis identifies Raf1 as a crucial mediator of BCR-ABL - Driven leukemogenesis

Albers, C., Illert, A. L., Miething, C., Leischner, H., Thiede, M., Peschel, C., Duyster, J. (August 2011) An RNAi-based system for loss-of-function analysis identifies Raf1 as a crucial mediator of BCR-ABL - Driven leukemogenesis. Blood, 118 (8). pp. 2200-2210. ISSN 00064971 (ISSN)

URL: https://www.ncbi.nlm.nih.gov/pubmed/21715303
DOI: 10.1182/blood-2010-10-309583

Abstract

Genetic loss-of-function studies in murine tumor models have been essential in the analysis of downstream mediators of oncogenic transformation. Unfortunately, these studies are frequently limited by the availability of genetically modified mouse strains. Here we describe a versatile method allowing the efficient expression of an oncogene and simultaneous knockdown of targets of interest (TOI) from a single retroviral vector. Both oncogene and TOI-specific miR30-based shRNA are under the control of the strong viral long terminal repeat promoter, resulting in a single shared RNA transcript. Using this vector in a murine syngeneic BM transplantation model for BCR-ABL - induced chronic myeloid leukemia, we find that oncogene expression andtargetknockdownin primary hematopoietic cells with this vector is efficient both in vitro and in vivo, and demonstrate that Raf1, but not BRAF, modulates BCR-ABL - dependent ERK activation and transformation of hematopoietic cells. This expression system could facilitate genetic loss-of-function studies and allow the rapid validation of potential drug targets in a broad range of oncogene-driven murine tumor models. © 2011 by The American Society of Hematology.

Item Type: Paper
Subjects: diseases & disorders > cancer
diseases & disorders > cancer > cancer types > leukemia
organism description > animal > mammal > rodent > mouse
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > shRNA
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > enzymes > kinase > tyrosine kinase
CSHL Authors:
Communities: CSHL labs > Lowe lab
Depositing User: CSHL Librarian
Date: August 2011
Date Deposited: 26 Oct 2011 19:29
Last Modified: 21 Feb 2018 21:38
Related URLs:
URI: http://repository.cshl.edu/id/eprint/15596

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