CBFA2T3 Antibody

What is CBFA2T3 and why is it important in hematological research?

CBFA2T3 (also known as MTG16 or ETO2) is a master transcriptional corepressor that plays critical roles in hematopoiesis. It functions by facilitating transcriptional repression through its association with DNA-binding transcription factors and recruitment of other corepressors and histone-modifying enzymes .

The significance of CBFA2T3 in hematological research stems from its:

• Role in maintaining leukemia stem cell (LSC) gene expression signatures

• Involvement in promoting relapse in acute myeloid leukemia (AML)

• Function in inhibiting terminal erythro-megakaryocytic differentiation

• Participation in maintaining stemness of long-term hematopoietic stem cells

• Formation of oncogenic fusion proteins such as CBFA2T3-GLIS2 and RUNX1-CBFA2T3 in various forms of AML

These characteristics make CBFA2T3 a critical target for researchers studying hematological malignancies and normal hematopoietic development.

What are the primary applications for CBFA2T3 antibodies in leukemia research?

CBFA2T3 antibodies serve multiple crucial research applications in the field of leukemia research:

• Mechanistic studies: Investigating how CBFA2T3 regulates gene expression through chromatin remodeling, particularly in the context of retinoic acid receptor (RAR) target gene repression

• Prognostic biomarker detection: Examining CBFA2T3 expression levels as a marker for leukemia stem cell signatures and potential relapse prediction

• Differentiation therapy research: Studying CBFA2T3's role in inhibiting ATRA-induced myeloid differentiation, which is particularly important for developing therapeutic strategies for AML subtypes resistant to differentiation therapy

• Fusion protein detection: Identifying CBFA2T3-GLIS2 and other fusion proteins in pediatric AML and rare leukemia subtypes

• Chromatin immunoprecipitation (ChIP): Mapping CBFA2T3 binding sites across the genome to understand its target genes and regulatory networks

These applications collectively contribute to our understanding of leukemogenesis and potential therapeutic interventions.

What experimental controls should be included when using CBFA2T3 antibodies?

When designing experiments with CBFA2T3 antibodies, the following controls are essential:

Positive controls:

• Cell lines with known CBFA2T3 expression (e.g., U937 AML cells)

• Recombinant CBFA2T3 protein for Western blot standardization

• CBFA2T3-overexpressing cells as a positive control reference

Negative controls:

• CBFA2T3 knockout or CRISPR/Cas9-edited cell lines (e.g., U937 C cells)

• Cell lines with minimal CBFA2T3 expression

• Secondary antibody-only controls for immunostaining applications

Validation controls:

• siRNA/shRNA-mediated CBFA2T3 knockdown to confirm antibody specificity

• Peptide competition assays to verify binding specificity

• Use of multiple antibodies targeting different epitopes of CBFA2T3

Normalization control:

• Housekeeping genes/proteins unaffected by CBFA2T3 manipulation (e.g., RPL30)

How should ChIP-Seq experiments with CBFA2T3 antibodies be optimized for studying its role in transcriptional regulation?

Optimizing ChIP-Seq experiments with CBFA2T3 antibodies requires several methodological considerations:

Antibody selection and validation:

• Use antibodies validated specifically for ChIP applications

• Confirm specificity through sequential ChIP or reciprocal IP with known interacting partners

• Perform ChIP-qPCR validation on known CBFA2T3 target regions before proceeding to sequencing

Crosslinking optimization:

• CBFA2T3 functions within multi-protein complexes, requiring careful crosslinking optimization

• Test both formaldehyde (1-2%) and dual crosslinking approaches (DSG followed by formaldehyde)

• Optimize crosslinking time (10-15 minutes) to preserve complex integrity without overfixation

Sonication parameters:

• Adjust sonication conditions to achieve 200-500bp fragments

• Verify fragmentation efficiency through gel electrophoresis

• Consider chromatin accessibility differences between cell types

Bioinformatic analysis approaches:

• Use specialized peak calling algorithms suitable for transcriptional corepressors

• Integrate with histone modification data (H3K27ac, H3K9ac) to identify active vs. repressed regions

• Perform de novo motif analysis to identify DNA-binding partners of CBFA2T3

• Analyze genomic distribution of binding sites (promoters, enhancers, etc.)

• Integrate with transcriptomic data to correlate binding with gene expression changes

Studies have shown CBFA2T3 preferentially targets myeloid-specific enhancers associated with terminal differentiation and overlaps significantly with RARα/RXRα cistromes in AML cells .

What are the optimal conditions for detecting CBFA2T3 by Western blotting in different cellular contexts?

Achieving optimal Western blot results for CBFA2T3 requires specific methodological adjustments based on cell type and context:

Lysis and extraction considerations:

• Use RIPA buffer supplemented with protease inhibitors for whole cell lysates

• Consider nuclear extraction methods as CBFA2T3 is primarily nuclear

• Include phosphatase inhibitors to preserve post-translational modifications

• Add deacetylase inhibitors (e.g., sodium butyrate) to preserve acetylation status

Protein separation parameters:

• Use 8-10% SDS-PAGE gels for optimal separation

• Expected molecular weight: 62-65 kDa observed (theoretical 71 kDa)

• Extended running time may improve resolution of post-translationally modified forms

Transfer and detection optimization:

• Semi-dry transfer at lower amperage (0.8 mA/cm²) for 60-90 minutes

• PVDF membranes preferable over nitrocellulose for CBFA2T3 detection

• Block with 5% BSA in TBST rather than milk to reduce background

• Primary antibody dilution: 1:500-1:2000 (optimize for each antibody)

• Extended primary antibody incubation (overnight at 4°C) improves sensitivity

Cell-type specific considerations:

• AML cell lines (U937, NB-4): Standard protocols generally effective

• Primary patient samples: Require gentler lysis and more concentrated lysates

• CBFA2T3-fusion expressing cells: May show aberrant migration patterns

Validation through multiple approaches including knockdown controls is essential to confirm specificity.

How can CBFA2T3 antibodies be effectively used to study its interaction with chromatin-modifying enzymes?

CBFA2T3 functions through interactions with various chromatin-modifying enzymes. Studying these interactions requires specialized methodological approaches:

Co-immunoprecipitation (Co-IP) optimization:

• Use gentle lysis buffers (150-200mM NaCl) to preserve protein complexes

• Include low concentrations of detergents (0.1-0.3% NP-40) to maintain interactions

• Add protease and phosphatase inhibitors to preserve complex integrity

• Consider crosslinking for transient interactions

• Reverse IP validation to confirm specificity of interactions

Sequential ChIP (Re-ChIP) approach:

• First immunoprecipitate with CBFA2T3 antibody

• Elute complexes under mild conditions

• Perform second ChIP with antibodies against histone acetyltransferases (p300, GCN5) or histone deacetylases (HDACs)

• This confirms co-occupancy at specific genomic loci

Proximity ligation assay (PLA):

• Use CBFA2T3 antibody in combination with antibodies against known partners (p300, GCN5, HDACs)

• Allows visualization of protein interactions in situ with subcellular resolution

• Quantify interaction signals in different cellular compartments

Mass spectrometry approaches:

• Immunoprecipitate CBFA2T3 complexes

• Perform tandem mass spectrometry to identify interacting partners

• Compare interaction profiles under different conditions (e.g., ATRA treatment)

Research has shown that CBFA2T3 competitively prevents p300 and GCN5 recruitment to chromatin, and these interactions are critical for its function in preventing myeloid differentiation in AML cells .

What are the common technical challenges when performing ChIP with CBFA2T3 antibodies and how can they be addressed?

Researchers often encounter several technical challenges when conducting ChIP experiments with CBFA2T3 antibodies:

Challenge 1: Low signal-to-noise ratio

• Cause: CBFA2T3 doesn't bind DNA directly but through protein complexes

• Solution:

  • Increase crosslinking time (15-20 minutes)

  • Use dual crosslinking with protein-protein crosslinkers like DSG followed by formaldehyde

  • Reduce sonication intensity to preserve protein complexes

  • Use higher antibody concentration (2-4 μg per reaction)

Challenge 2: Epitope masking

• Cause: Protein interactions may block antibody binding sites

• Solution:

  • Test multiple antibodies targeting different CBFA2T3 epitopes

  • Adjust crosslinking conditions to find optimal preservation without masking

  • Consider native ChIP approaches for certain applications

Challenge 3: Cell type-specific binding patterns

• Cause: CBFA2T3 cistrome varies significantly between cell types

  • In U937 AML cells, CBFA2T3 predominantly targets RARα/RXRα binding sites

  • In other cell types, binding patterns may differ substantially

• Solution:

  • Perform broad peak calling with less stringent thresholds

  • Validate binding with ChIP-qPCR on multiple targets

  • Use cell type-appropriate positive controls

Challenge 4: Data analysis complexities

• Cause: CBFA2T3 binding may not correlate directly with gene expression changes

• Solution:

  • Integrate ChIP-seq with RNA-seq data from matched samples

  • Analyze chromatin accessibility (ATAC-seq) in parallel

  • Examine histone modifications to contextualize binding sites

A systematic approach to optimization with appropriate controls is essential for successful CBFA2T3 ChIP experiments.

How can non-specific binding be distinguished from true CBFA2T3 signals in immunostaining applications?

Distinguishing specific from non-specific staining is crucial for accurate CBFA2T3 immunostaining:

Antibody validation strategies:

• Genetic validation: Compare staining in CBFA2T3 knockout/knockdown cells vs. controls

• Peptide competition: Pre-incubate antibody with immunizing peptide to block specific binding

• Multiple antibody validation: Confirm staining pattern with independent antibodies targeting different epitopes

• Isotype controls: Use matched isotype antibodies at equivalent concentrations

Technical optimization approaches:

• Blocking optimization:

  • Extended blocking (2+ hours) with 5-10% normal serum

  • Addition of 0.1-0.3% Triton X-100 to reduce hydrophobic interactions

  • Consider specialized blocking agents (e.g., Image-iT FX Signal Enhancer)

• Antibody titration:

  • Test serial dilutions to identify optimal concentration

  • Monitor signal-to-noise ratio across dilution series

  • Use quantitative image analysis to determine optimal concentration

• Signal verification through orthogonal approaches:

  • Correlate immunostaining with fluorescent protein tagging

  • Verify localization pattern with cellular fractionation and Western blotting

  • Use RNA in situ hybridization to correlate with protein expression

True CBFA2T3 signal characteristics:

• Predominantly nuclear localization with nucleolar exclusion

• Intensity correlates with known expression patterns (high in stem/progenitor cells, reduced upon differentiation)

• Signal abolished by specific competition or in knockout cells

• Co-localization with known interaction partners

Implementing these strategies systematically will help distinguish true CBFA2T3 signals from artifacts.

What strategies can address the challenge of detecting CBFA2T3-fusion proteins in patient samples?

Detecting CBFA2T3 fusion proteins in patient samples presents unique challenges requiring specialized approaches:

Sample preparation considerations:

• Fresh or flash-frozen samples preferred over FFPE for protein detection

• Gentle thawing procedures to minimize protein degradation

• Nuclear extraction methods to concentrate fusion proteins

• Consider density gradient separation to enrich blast populations in heterogeneous samples

Technical detection strategies:

• Western blotting optimization:

  • Use gradient gels (4-12%) to better resolve high molecular weight fusion proteins

  • Extended transfer times (overnight at low voltage) for large fusion proteins

  • Dual antibody approach: probe with both CBFA2T3 and fusion partner antibodies

  • Expected molecular weights:

    • CBFA2T3-GLIS2: ~110-120 kDa

    • RUNX1-CBFA2T3: ~140 kDa

• Immunofluorescence approaches:

  • Co-staining with antibodies against both fusion partners

  • Use proximity ligation assay (PLA) to detect fusion-specific protein juxtapositions

  • Optimized antigen retrieval protocols for clinical samples

• Flow cytometry applications:

  • Combined surface and intracellular staining protocols

  • Include markers for blast identification (CD34, CD117)

  • Gating on relevant populations to increase sensitivity

Complementary molecular techniques:

• RT-PCR or RNA-seq to confirm fusion at transcript level

• FISH to detect chromosomal rearrangements

• Consider specialized techniques like NanoString for targeted fusion detection

Validation in model systems:

• Use cell lines with known CBFA2T3 fusions as positive controls

• Compare detection limits across methodologies

• Consider synthetic fusion protein standards for quantification

The challenge of detecting rare CBFA2T3 fusions requires a multi-modal approach combining protein and nucleic acid-based methods for maximal sensitivity and specificity .

How can researchers distinguish between the roles of wild-type CBFA2T3 and fusion proteins in leukemogenesis?

Distinguishing the functions of wild-type CBFA2T3 versus fusion proteins requires systematic experimental approaches:

Differential detection strategies:

• Use domain-specific antibodies that selectively recognize:

  • N-terminal regions (present in both wild-type and C-terminal fusions)

  • C-terminal regions (absent in CBFA2T3-GLIS2 but present in wild-type)

  • Fusion-specific epitopes at junction points

• Implement size-based separation techniques to distinguish proteins based on molecular weight differences

Functional characterization approaches:

• Selective knockdown/knockout experiments:

  • siRNA targeting fusion-specific junctions

  • CRISPR-Cas9 strategies with guide RNAs specific to fusion or wild-type sequences

  • Rescue experiments with selective re-expression

• Domain-specific function analysis:

  • Create domain deletion constructs to identify critical functional regions

  • Compare chromatin binding profiles of wild-type vs. fusion proteins

  • Assess differential protein interaction partners

• Transcriptional impact analysis:

  • Wild-type CBFA2T3 represses retinoic acid receptor target genes

  • CBFA2T3-GLIS2 fusion activates BMP/GATA target genes

  • Compare gene expression signatures through RNA-seq:

Patient sample analysis approaches:

• Correlate fusion status with clinical parameters and outcomes

• Compare bone marrow morphology and immunophenotype

• Evaluate resistance to differentiation therapy in wild-type vs. fusion-expressing samples

These complementary approaches allow researchers to delineate the specific contributions of wild-type CBFA2T3 and its fusion derivatives to leukemogenesis.

What approaches can be used to study the dynamics of CBFA2T3 regulation during ATRA-induced differentiation?

Studying CBFA2T3 dynamics during ATRA-induced differentiation requires temporal analysis approaches:

Time-course experimental design:

• Sample collection at multiple timepoints (0h, 6h, 24h, 48h, 72h post-ATRA treatment)

• Use standardized ATRA concentrations (1-2 μM) for consistency

• Include vehicle controls at each timepoint

• Consider parallel treatment with ATRA + CBFA2T3 knockdown/overexpression

Multi-omics analysis approaches:

• Protein level dynamics:

  • Western blotting for total CBFA2T3 protein levels

  • Phospho-specific antibodies to track post-translational modifications

  • Subcellular fractionation to monitor nuclear/cytoplasmic distribution

  • Proteasome inhibition studies to assess degradation kinetics

  • Research shows ATRA treatment rapidly down-regulates CBFA2T3 in NB-4 cells

• Genomic binding dynamics:

  • ChIP-seq at multiple timepoints to track chromatin occupancy changes

  • Focus on RARα target regions showing dynamic regulation

  • Integrate with chromatin accessibility (ATAC-seq) data

  • Monitor displacement of CBFA2T3 from target loci during differentiation

• Transcriptional regulation analysis:

  • RNA-seq to correlate CBFA2T3 binding with gene expression changes

  • Nascent RNA analysis (e.g., PRO-seq) to detect immediate transcriptional effects

  • GRO-seq to monitor changes in transcriptional elongation

Mechanistic interrogation approaches:

• Assess changes in CBFA2T3 interaction partners during ATRA treatment

• Monitor recruitment kinetics of histone modifiers (p300, GCN5, HDACs)

• Measure histone acetylation dynamics at CBFA2T3 target sites

• Correlate molecular changes with morphological differentiation markers

This multi-faceted approach enables comprehensive understanding of CBFA2T3's role in ATRA response resistance, potentially informing therapeutic strategies for differentiation-resistant AML .

How can CBFA2T3 antibody-based techniques contribute to understanding therapy resistance mechanisms in AML?

CBFA2T3 antibody-based techniques provide critical insights into AML therapy resistance mechanisms:

Patient sample analysis approaches:

• Expression level assessment:

  • Compare CBFA2T3 protein levels between diagnosis and relapse samples

  • Correlate expression with clinical outcomes and response to differentiation therapy

  • Analyze subcellular localization patterns in responsive vs. resistant samples

  • Studies show CBFA2T3 maintains leukemia stem cell gene expression and promotes relapse

• ChIP-seq profiling in paired samples:

  • Compare CBFA2T3 genomic binding before and after therapy

  • Identify sites with persistent binding in resistant cases

  • Correlate with changes in chromatin structure and gene expression

  • Focus on genes mediating stemness and self-renewal properties

Functional validation studies:

• Experimental modulation approaches:

  • Use CRISPR/Cas9 to create CBFA2T3 knockout models

  • Assess sensitivity to ATRA and other differentiation agents

  • Re-express CBFA2T3 to confirm causal relationship to resistance

  • Test combinatorial approaches targeting CBFA2T3 and differentiation pathways

• Pathway analysis studies:

  • Examine CBFA2T3 effects on RAR-dependent gene transcription

  • Monitor histone acetyltransferase recruitment and histone acetylation

  • Assess chromatin accessibility changes at therapy-resistance genes

Translational research applications:

• Develop CBFA2T3 expression as a predictive biomarker for ATRA response

• Identify patients who might benefit from combination therapy targeting CBFA2T3

• Design rational drug combinations based on CBFA2T3 mechanistic insights

Research has demonstrated that loss of CBFA2T3 sensitizes AML cells to ATRA-induced differentiation, with CBFA2T3 knockdown cells showing a nearly 3-fold reduction in ED50 for ATRA response . The differential ability of AML subtypes to repress CBFA2T3 correlates with clinical outcomes - t(8;21) AML naturally represses CBFA2T3 through RUNX1-RUNX1T1 and has better prognosis, while other AML subtypes lacking this repression mechanism show higher relapse rates .

What is the significance of CBFA2T3 isoforms in hematological research and how can they be differentially detected?

CBFA2T3 exists in multiple isoforms with distinct functional implications for hematological research:

Major CBFA2T3 isoforms:

• CBFA2T3 isoform 1 (MTG16a): Full-length protein (653 amino acids)

• CBFA2T3 isoform 2 (MTG16b): Shorter isoform with alternative N-terminus

• CBFA2T3 isoform 3: Subject to alternative splicing affecting functional domains

• Post-translationally modified forms: Various phosphorylation, acetylation states

Functional differences between isoforms:

• Isoform 2 functions as an A-kinase-anchoring protein

• Different isoforms show distinct interaction patterns with transcription factors

• Varying repressive capacities against specific target genes

• Differential subcellular localization patterns

Isoform-specific detection approaches:

• Antibody-based strategies:

  • Use isoform-specific antibodies targeting unique regions

  • Western blotting can reveal distinct molecular weight bands:

    • Isoform 1: ~62-65 kDa (observed)

    • Isoform 2: Slightly lower molecular weight

  • Validate specificity with isoform-specific knockdown experiments

• mRNA analysis methods:

  • Design qPCR primers spanning isoform-specific exon junctions

  • Use RNA-seq with computational pipelines designed to detect splice variants

  • Consider targeted approaches like NanoString for quantitative isoform profiling

Research applications for isoform analysis:

• Compare isoform expression ratios across differentiation stages

• Assess whether specific isoforms correlate with treatment response

• Determine if certain AML subtypes preferentially express specific isoforms

• Investigate isoform-specific protein interactions that may impact leukemogenesis

Methodological considerations:

• Ensure antibodies can distinguish between closely related isoforms

• Consider using epitope-tagged constructs for overexpression studies

• Implement mass spectrometry approaches for unambiguous isoform identification

• Design isoform-specific genetic manipulation strategies

Understanding the specific roles of CBFA2T3 isoforms may reveal more nuanced approaches to targeting this protein in leukemia therapy and provide insights into the differential functions of CBFA2T3 in normal and malignant hematopoiesis.

How might single-cell approaches with CBFA2T3 antibodies reveal new insights about heterogeneity in AML?

Single-cell technologies combined with CBFA2T3 antibodies offer unprecedented opportunities to investigate AML heterogeneity:

Single-cell protein analysis approaches:

• Mass cytometry (CyTOF):

  • Incorporate CBFA2T3 antibodies into panels with surface markers and other transcription factors

  • Create comprehensive differentiation trajectories based on protein expression

  • Identify rare subpopulations with distinct CBFA2T3 expression patterns

  • Correlate with functional markers of stemness and resistance

• Single-cell Western blotting:

  • Quantify CBFA2T3 protein levels in individual cells

  • Correlate with differentiation markers at the single-cell level

  • Identify cells with aberrant CBFA2T3 expression or post-translational modifications

• Imaging mass cytometry:

  • Visualize CBFA2T3 expression in spatial context within bone marrow architecture

  • Correlate with microenvironmental factors that may influence expression

  • Detect rare cells with nuclear CBFA2T3 localization patterns

Integrated multi-omics approaches:

• CITE-seq with CBFA2T3 antibodies:

  • Simultaneously profile surface protein markers and transcript expression

  • Correlate CBFA2T3 protein levels with target gene expression

  • Create high-dimensional maps of cellular states in AML

• Single-cell CUT&Tag:

  • Map CBFA2T3 chromatin binding at single-cell resolution

  • Identify cell-specific regulatory regions

  • Correlate binding patterns with differentiation trajectories

Potential research insights:

• Identification of rare therapy-resistant cells with distinct CBFA2T3 profiles

• Discovery of new cellular subpopulations with unique dependencies

• Understanding the dynamics of CBFA2T3 expression during disease progression and treatment

• Correlation of CBFA2T3 status with leukemia stem cell properties at single-cell resolution

These approaches could reveal why certain AML cells within a heterogeneous population survive therapy and contribute to relapse, potentially identifying new therapeutic vulnerabilities related to CBFA2T3 function .

What are the emerging strategies for targeting CBFA2T3 function in AML therapy development?

Recent research suggests several promising strategies for targeting CBFA2T3 in AML therapy:

Direct targeting approaches:

• Protein degradation technologies:

  • Development of PROTACs (Proteolysis Targeting Chimeras) against CBFA2T3

  • Molecular glue degraders exploiting ubiquitin-proteasome machinery

  • These approaches would mimic the natural down-regulation of CBFA2T3 seen during ATRA-induced differentiation

• Domain-specific inhibitors:

  • Small molecules targeting the MYND domain crucial for protein-protein interactions

  • Disruption of NHR domains involved in oligomerization

  • Peptide-based inhibitors of specific interaction interfaces

Functional pathway modulation:

• Epigenetic sensitization:

  • HDAC inhibitors to counteract CBFA2T3's repressive functions

  • Bromodomain inhibitors to modulate acetylation-dependent interactions

  • Combination with ATRA to enhance differentiation responses

• Transcriptional regulation:

  • Compounds that promote CBFA2T3 transcriptional downregulation

  • Targeting the NM_005187 promoter activation by GCN5

  • RNA-based therapeutics to reduce CBFA2T3 expression

Combination therapy approaches:

• ATRA plus CBFA2T3 inhibition for non-APL AML subtypes

• Targeting CBFA2T3 in combination with conventional chemotherapy

• Sequential therapy approaches for maximal differentiation induction

Biomarker-driven strategies:

• CBFA2T3 expression levels as predictive biomarkers for therapy selection

• Patient stratification based on CBFA2T3-dependent gene signatures

• Monitoring CBFA2T3 status during treatment as a response indicator

Research has demonstrated that CBFA2T3 inhibition sensitizes normally resistant AML cells to ATRA-induced differentiation, suggesting that targeting CBFA2T3 could expand the utility of differentiation therapy beyond acute promyelocytic leukemia to other AML subtypes .

How can CBFA2T3 antibodies contribute to understanding the intersection between normal hematopoiesis and leukemogenesis?

CBFA2T3 antibodies provide valuable tools for investigating the critical intersection between normal hematopoietic development and leukemic transformation:

Comparative analysis approaches:

• Developmental stage-specific profiling:

  • Map CBFA2T3 expression across the hematopoietic hierarchy

  • Compare chromatin binding patterns in normal HSCs vs. leukemic stem cells

  • Identify stage-specific cofactors and target genes

  • Research shows CBFA2T3 maintains stemness in long-term HSCs and expands leukemia stem cells

• Lineage commitment studies:

  • Track CBFA2T3 dynamics during normal myeloid differentiation

  • Compare with aberrant patterns in leukemic differentiation block

  • Investigate how CBFA2T3 inhibits terminal erythro-megakaryocytic differentiation in normal and malignant contexts

• Microenvironmental interaction analysis:

  • Examine how niche factors regulate CBFA2T3 expression

  • Compare responses to differentiation signals between normal and leukemic cells

  • Investigate cross-talk with other key hematopoietic regulators

Functional perturbation approaches:

• Loss-of-function models:

  • Conditional knockout in normal hematopoietic stem/progenitor cells

  • Compare phenotypes with CBFA2T3 knockdown in leukemic cells

  • Time-resolved analysis of differentiation trajectories

• Gain-of-function studies:

  • Overexpression in defined progenitor populations

  • Assessment of self-renewal and differentiation capacity

  • Evaluation of transformation potential and cooperating mutations

Translational implications:

• Identification of CBFA2T3-dependent vulnerabilities specific to leukemic cells

• Development of therapeutic strategies that selectively target leukemic stem cells

• Understanding mechanism of relapse through CBFA2T3-mediated gene programs