Rapid generation of a tetracycline-inducible BCR-ABL defective retrovirus using a single autoregulatory retroviral cassette

  title={Rapid generation of a tetracycline-inducible BCR-ABL defective retrovirus using a single autoregulatory retroviral cassette},
  author={Aymeric Dugray and JF Geay and Adlen Foudi and M L Bonnet and William Vainchenker and Françoise Wendling and Fawzia Louache and Ag Turhan},
The development of chronic myelogenous leukemia (CML) models in mice using an inducible BCR-ABL gene has been hampered by the requirement of sequential expression of tTA (Tet repressor-VP16 fusion protein) and Tet-OP sequences in the same cells after separate transfection. This double transfection strategy is time consuming as it requires screening of many hundreds of individual clones and cannot be applied to primary hematopoietic cells. To generate a tetracycline-inducible BCR-ABL retrovirus… 

Down-regulation of BRCA1 in BCR-ABL-expressing hematopoietic cells.

It is reported that the expression of the p210 BCR-ABL fusion protein leads to a down-regulation of BRCA1 protein, a gene product involved in the maintenance of genome integrity, which reveals a novel link between the oncoprotein BCR -ABL and the tumor-suppressor protein BRC a1.

Tight regulation from a single tet-off rAAV vector as demonstrated by flow cytometry and quantitative, real-time PCR

Results demonstrate that exceedingly tight regulation of transgene expression is possible using the tet-off system in the context of a self-regulating rAAV vector and that the specific orientation of two promoters relative to each other and to the ITRs is important.

Chronic myeloid leukemia stem cells possess multiple unique features of resistance to BCR-ABL targeted therapies

It is shown that the lesser effect of imatinib mesylate on the 3-week output of cells produced in vitro from lin−CD34+CD38− CML (stem) cells compared with cultures initiated with the CD38+ subset of lin− CD34+ cells is markedly enhanced when conditions of reduced growth factor stimulation are used.

Strategies of Conditional Gene Expression in Myocardium

This chapter outlines the packaging of myocardium-specific inducible expression systems into lentiviral vectors, in which a transgene and a reporter gene are transduced into cardiomyocytes and can be monitored/tracked with bioluminescence imaging (BLI) and positron emission tomography (PET).

AHI-1 interacts with BCR-ABL and modulates BCR-ABL transforming activity and imatinib response of CML stem/progenitor cells

It is demonstrated that overexpression of Ahi-1/AHI-1 in murine and human hematopoietic cells confer growth advantages in vitro and induce leukemia in vivo, enhancing effects of BCR-ABL.

Osteopontin is upregulated by BCR-ABL.

ENOX2 NADH Oxidase: A BCR-ABL1-dependent cell surface and secreted redox protein in chronic myeloid leukemia (CML)

The results demonstrate for the first time the upregulation of a secreted Redox protein in a BCR-ABL1-dependent manner in CML and suggest that ENOX2 (through its transcriptional program) plays a significant role in the B CR-ABl1 leukemogenesis.

Targeting Primitive Chronic Myeloid Leukemia Cells by Effective Inhibition of a New AHI-1–BCR-ABL–JAK2 Complex

Simultaneously targeting BCR-ABL and JAK2 activities in C ML stem/progenitor cells may improve outcomes in patients destined to develop IM resistance, and was effective against treatment-naive CML stem cells from patients who subsequently proved to be resistant to IM therapy.

An optimized conditional suicide switch using doxycycline-dependent expression of human tBid

Human tBid, an endogenous effector protein with low immunogenic potential, and doxycycline, an inducer with low cytotoxicity and a long record of safe use in humans, provides a tool for various applications, e.g. adoptive immune transfer.

p210BCR-ABL inhibits SDF-1 chemotactic response via alteration of CXCR4 signaling and down-regulation of CXCR4 expression.

It is reported here that the migratory response to SDF-1 was profoundly altered in blast crisis, whereas chronic-phase CD34(+) cells migrated normally to this chemokine, indicating that p210(BCR-ABL) could affect CXCR4 by more than one mechanism and suggest that down-regulation of CX CR4 may have important implications in chronic myelogenous leukemia pathogenesis.



Rapid retroviral delivery of tetracycline-inducible genes in a single autoregulatory cassette.

The current cassette of the retroviral construct (SIN-RetroTet vector) allows rapid delivery of inducible genes and should have broad applications to cultured cells, transgenic animals, and gene therapy.

A stable human-derived packaging cell line for production of high titer retrovirus/vesicular stomatitis virus G pseudotypes.

We have generated a human 293-derived retroviral packaging cell line (293GPG) capable of producing high titers of recombinant Moloney murine leukemia virus particles that have incorporated the

Bcr-Abl efficiently induces a myeloproliferative disease and production of excess interleukin-3 and granulocyte-macrophage colony-stimulating factor in mice: a novel model for chronic myelogenous leukemia.

It is demonstrated that expression of Bcr-Abl can induce a CML-like leukemia in mice much more efficiently and reproducibly than in previously reported mouse CML models, probably due to efficient expression in the correct target cell(s).

Secondary mutation maintains the transformed state in BaF3 cells with inducible BCR/ABL expression.

The tight conditional expression of BCR/ABL in the TonB210.1 cell line affords the opportunity to study several interesting aspects of the biology of B CR/ABl, including activation of critical signaling pathways and transcriptional programs, and its potential role in genomic instability.

Efficient and Rapid Induction of a Chronic Myelogenous Leukemia-Like Myeloproliferative Disease in Mice Receiving P210 bcr/abl-Transduced Bone Marrow

Results demonstrate that murine CML recapitulates important features of human CML and should be an excellent model for addressing specific issues relating to the pathogenesis and treatment of this disease.

Tight control of gene expression in mammalian cells by tetracycline-responsive promoters.

  • M. GossenH. Bujard
  • Biology
    Proceedings of the National Academy of Sciences of the United States of America
  • 1992
Control elements of the tetracycline-resistance operon encoded in Tn10 of Escherichia coli have been utilized to establish a highly efficient regulatory system in mammalian cells that is suitable for creation of "on/off" situations for such genes in a reversible way.

BCR-ABL and constitutively active erythropoietin receptor (cEpoR) activate distinct mechanisms for growth factor-independence and inhibition of apoptosis in Ba/F3 cell line

The interleukin-3 dependent murine Ba/F3 cell line has been widely used as an experimental model of cell transformation by BCR – ABL oncogenes as assessed by induction of growth-factor-independence

Tet B or not tet B: advances in tetracycline-inducible gene expression.

  • H. BlauF. Rossi
  • Biology
    Proceedings of the National Academy of Sciences of the United States of America
  • 1999
Modulating gene expression in cycles that mimic endogenous patterns is highly desirable, and avoiding toxic levels is a must, for gene therapy, regulation is crucial.

Induction of chronic myelogenous leukemia in mice by the P210bcr/abl gene of the Philadelphia chromosome.

It is demonstrated that P210bcr/abl expression can induce chronic myelogenous leukemia and retrovirus-mediated expression of the protein provides a murine model system for further analysis of the disease.

Vesicular stomatitis virus G pseudotyped retrovector mediates effective in vivo suicide gene delivery in experimental brain cancer.

Direct in vivo tumor-targeting with "suicide" viral vectors is limited by either inefficient gene transfer (i.e., retroviral vectors) or indiscriminate transfer of a conditionally toxic gene to