Our scientists have years of experience at performing gene editing with CRISPR/Cas9, from designing gRNA to transfection and single clone generation. Creative Biogene is offering a series of knockout cell lines developed by CRISPR/Cas9 system. These cell lines provide you with a convenient means to study gene functions

Marcia Brady

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BCR-ABL, or BCR-ABL1, is a fusion gene formed by the combination of two genes, known as BCRand ABL. BCR-ABL fusion gene is found in certain types of leukemia, for example, in almost allpatients bearing chronic myeloid leukemia (CML), in some patients with acute lymphoblasticleukemia (ALL) and rarely in patients with acute myelogenous leukemia (AML).Figure 1. Formation of BCR-ABL fusion geneA series of specific small-moleculeinhibitors have been designed to inhibitthe tyrosine kinase activity of the Bcr-Abloncoprotein, including Imatinib,Nilotinib and Dasatinib. Some of whichhave been approved for use in clinical,and several of which are in clinical trials.T315I mutationThe T315I is a missense mutation resulting in anamino acid substitution of BCR-ABL fromthreonine (T) to isoleucine (I) at position 315.T315I substitution leads to the absence of acritical hydrogen bond between imatinib andthe ABL kinase, and alters the three-dimensionalstructure of the kinase protein binding site.Several studies have shown that presence ofT315I in BCR-ABL1 mediates resistance toImatinib and many TKIs except Ponatinib.Figure 2. BCR/ABL kinase inhibition by imatinibTel: 1-631-626-9181    |    Fax: 1-631-614-7828    |    Email: info@creative-biogene.com |    45-1 Ramsey Road Shirley, NY 11967 USA    |    www.creative-biogene.com BaF3-BCR-ABL & BaF3-BCR-ABL-T315I Stable Cell Lines - In Vitro Screening Tool for CML DrugsFigure 5. Kinase inhibitor analysis using ourBaF3-BCR-ABL cell line. The IC50 of Imatinib,Ponatinib and Dasatinib were 4.54 μM, 37 nMand 11 nM respectively.Figure 6. Kinase inhibitor analysis using our BaF3-BCR-ABL-T315I cell line. The IC50 of DasatinibImatinib and Ponatinib were >10 μM, >10 μM and45 nM respectively.Application1. Cell-based screening of BCR-ABL tyrosine-kinase inhibitors (TKI)2. Cell viability assay3. In vivo efficacy studyReferences1. Weisberg E1, Manley PW, Cowan-Jacob SW,Hochhaus A, Griffin JD. Second generation inhibitorsof BCR-ABL for the treatment of imatinib-resistantchronic myeloid leukaemia. Nat Rev Cancer. 2007May;7(5):345-56.2. Daigo A, Tetsuzo T, Seiichi O, Kousuke N, Kazuma O,Activity of a novel Aurora kinase inhibitor against theT315I mutant form of BCR ‐ ABL: In vitro and in vivostudies. Cancer Science. 2008 June; 99 (6):1251–1257.Figure 4. WB analysis of BCR-ABL-T315I expressionFigure 3. WB analysis of BCR-ABL expressionTel: 1-631-626-9181    |    Fax: 1-631-614-7828    |    Email: info@creative-biogene.com |    45-1 Ramsey Road Shirley, NY 11967 USA    |    www.creative-biogene.com Creative Biogene has developed BCR-ABL Stable CellLine - BaF3 [CSC-RO0124] and BCR-ABL-T315I StableCell Line - BaF3 [CSC-RO0125] which are suitable foruse in cell-based in vitro screening of drugcandidatesagainst chronic myeloid leukemia.Why Using BaF3 Cell Line?Ba/F3 is a murine hematopoietic cell line which isestablished from bone marrow of BALB/c mouse. IL-3 isessential for Ba/F3 cell maintenance. Several advantageshave made this cell line to be a popular system in kinasedrug discovery.1. BaF3 grows in a fast speed which enables rapid experimental timeline;2. BaF3 is suitable as a transfection or transduction host;3. BaF3 becomes IL-3 independence when it constitutively expresses active tyrosine kinase through cell engineering.Our BaF3-BCR-ABL and BaF3-BCR-ABL-T315I stable celllines are constructed by stable delivery of exogenoushuman BCR-ABL, BCR-ABL-T315I encoding sequencesinto BaF3 cell genomes which are mediated byretrovirus transduction respectively. Western blotanalysis shows significant overexpression level of BCR-ABL, BCR-ABL-T315I in our cell lines. We have alsotested the IC 50 of BCR-ABL known inhibitors includingImatinib, Ponatinib and Dasatinib.

Stable Cell Lines

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Stable Cell Lines

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