Recombinant Didelphis marsupialis virginiana Bladder cancer-associated protein (BLCAP)

What is BLCAP and why is it significant in cancer research?

BLCAP (Bladder Cancer-Associated Protein) is a small 87-amino acid evolutionarily conserved protein with no homology to any known protein. It was originally identified from human bladder carcinoma and is considered a novel candidate tumor suppressor gene. The significance of BLCAP in cancer research stems from observations that loss of BLCAP mRNA expression correlates with the invasive potential of urothelial carcinomas (UCs), and differential expression has been noted across various cancer types including cervical, renal, human tongue carcinoma, and osteosarcoma . Further interest arises from evidence that BLCAP protein expression is lost during tumor progression, yet is overexpressed in approximately 20% of cases, where this overexpression correlates with poor survival, suggesting potential prognostic value .

What are the structural characteristics of recombinant Didelphis marsupialis virginiana BLCAP?

Recombinant Didelphis marsupialis virginiana (North American opossum) BLCAP is a full-length protein consisting of 87 amino acids (positions 1-87). The amino acid sequence is MYCLQWLLPVLLIPKPLNPALWFSHSMFMGFYLLSFLLERKPCTICALVFLAALFLICYSC WGNCFLYHCTGSHLPESAHDPRIVGT. When produced as a recombinant protein, it is typically tagged with a histidine tag (His-tag) at the N-terminus to facilitate purification. The protein is expressed in E. coli expression systems and usually achieves a purity greater than 90% as determined by SDS-PAGE . The structure features a highly conserved amino terminus that can be modified through RNA editing, potentially creating alternative protein isoforms with altered functions .

How does BLCAP expression differ between normal and cancerous tissues?

BLCAP exhibits differential expression patterns between normal and cancerous tissues. In normal tissues, BLCAP is ubiquitously expressed but with varying levels of RNA editing across different tissue types. For instance, heart and fibroblast tissues show low levels of RNA editing (5.1% at the Y/C site, 3.8% at the Q/R site, and 1.3% at the K/R site in heart tissue), while bladder tissue demonstrates higher editing activity (27.6% at the Y/C site, 15.8% at the Q/R site, and 5.3% at the K/R site) .

In cancerous tissues, there is a general downregulation of both BLCAP expression and RNA editing. For example, in white matter of normal brain tissue, editing activity at the Q/R site was 19.4%, which reduced to 0-4% in cancer tissue and cell lines. Similarly, the K/R site in white matter was edited to 19.4% but decreased to 0-4.2% in tumors and cancer cell lines . A correlation has been observed between decreased editing levels at Q/R and K/R sites and increased histological grade of malignancy in pediatric astrocytomas, suggesting potential diagnostic or prognostic applications .

What are the recommended protocols for reconstitution and storage of recombinant BLCAP protein?

For optimal handling of recombinant BLCAP protein, follow these methodological steps:

• Initial Handling: Briefly centrifuge the vial prior to opening to bring contents to the bottom.

• Reconstitution: Reconstitute the lyophilized protein in deionized sterile water to achieve a concentration of 0.1-1.0 mg/mL.

• Stabilization: Add glycerol to a final concentration of 5-50% (50% is the standard recommendation). This helps maintain protein stability during freeze-thaw cycles.

• Aliquoting: Divide the reconstituted protein into small working aliquots to minimize repeated freeze-thaw cycles.

• Storage Conditions:

  • Short-term working aliquots: Store at 4°C for up to one week

  • Long-term storage: Store at -20°C/-80°C

  • Avoid repeated freeze-thaw cycles as this can compromise protein integrity

• Buffer Information: The protein is typically provided in Tris/PBS-based buffer with 6% Trehalose at pH 8.0 .

This protocol maximizes protein stability and functionality for experimental applications while minimizing degradation.

What experimental approaches can be used to study the tumor suppressive function of BLCAP?

To investigate BLCAP's tumor suppressive functions, researchers should consider these methodological approaches:

• Overexpression Studies: Transfect cancer cell lines with BLCAP expression vectors and assess:

  • Cell proliferation rates (using MTT/XTT assays or cell counting)

  • Apoptosis induction (using Annexin V/PI staining and flow cytometry)

  • Cell cycle analysis (using PI staining and flow cytometry)

  • Colony formation ability (using soft agar assays)

• Knockdown/Knockout Studies: Use siRNA, shRNA, or CRISPR-Cas9 to reduce or eliminate BLCAP expression in normal or low-grade cancer cells, then evaluate:

  • Changes in proliferation and invasive potential

  • Alterations in apoptotic resistance

  • Effects on tumor formation in animal models

• RNA Editing Analysis: Examine editing patterns of BLCAP transcripts in normal versus cancerous tissues using:

  • RT-PCR followed by direct sequencing of the amplicons

  • Analysis of editing sites (Y/C, Q/R, and K/R) to correlate with malignancy grades

• Protein-Protein Interaction Studies: Identify BLCAP-interacting partners through:

  • Co-immunoprecipitation experiments

  • Yeast two-hybrid screening

  • Mass spectrometry-based interactome analysis

• Functional Pathway Analysis: Investigate how BLCAP affects known cancer signaling pathways through:

  • Western blot analysis of key pathway proteins

  • Transcriptome analysis using RNA-seq

  • Phosphoproteome analysis

These approaches provide complementary data on the molecular mechanisms underlying BLCAP's tumor suppressive roles, particularly relating to its capacity to inhibit cell growth and induce apoptosis as observed in cervical cancer HeLa cells and tongue carcinoma Tca8113 cell lines .

How can researchers accurately measure BLCAP protein expression levels in tissue samples?

For accurate quantification of BLCAP protein expression in tissue samples, researchers should implement a multi-faceted approach:

• Immunohistochemistry (IHC):

  • Use validated anti-BLCAP antibodies with appropriate controls

  • Employ tissue microarrays for high-throughput analysis

  • Implement standardized scoring systems (e.g., H-score or Allred) for semi-quantitative assessment

  • Consider automated image analysis software for objective quantification

• Western Blotting:

  • Extract proteins using optimized lysis buffers compatible with membrane proteins

  • Normalize protein loading using housekeeping proteins (β-actin, GAPDH)

  • Use chemiluminescence detection with standard curves for quantification

  • Consider multiplexed detection systems for simultaneous analysis of multiple proteins

• ELISA:

  • Develop sandwich ELISA assays using capture and detection antibodies specific for BLCAP

  • Include recombinant BLCAP protein standards for calibration curves

  • Validate assay linearity, sensitivity, and specificity

• Mass Spectrometry:

  • Implement targeted proteomics approaches such as Selected Reaction Monitoring (SRM) or Parallel Reaction Monitoring (PRM)

  • Use stable isotope-labeled peptide standards for absolute quantification

  • Focus on unique peptides that distinguish between edited and unedited BLCAP isoforms

• Normalization and Quality Control:

  • Always include appropriate positive and negative controls

  • Normalize to tissue area, cell number, or total protein content

  • Consider the use of multiple methodologies for cross-validation

This comprehensive approach allows for robust quantification of BLCAP protein levels and can reveal differences between normal and cancerous tissues, potentially serving as a prognostic biomarker in bladder cancer and other malignancies .

What is the significance of RNA editing in BLCAP transcripts?

RNA editing represents a critical post-transcriptional modification of BLCAP with substantial biological implications:

• Protein Isoform Generation: A-to-I editing events in BLCAP transcripts create alternative protein isoforms by changing genetically coded amino acids, particularly in the highly conserved amino terminus. The most significant editing sites include:

  • Y/C site: Converts tyrosine to cysteine

  • Q/R site: Changes glutamine to arginine

  • K/R site: Transforms lysine to arginine

• Tissue-Specific Regulation: Different tissues display distinctive ratios of edited and unedited BLCAP transcripts, suggesting tissue-specific regulation mechanisms. For example, bladder tissue shows higher editing rates (27.6% at Y/C site) compared to heart tissue (5.1% at Y/C site) .

• Cancer Association: A general decrease in BLCAP-editing levels is observed in astrocytomas, bladder cancer, and colorectal cancer compared to related normal tissues. This suggests that editing status may serve as a potential diagnostic tool or biomarker for distinguishing malignancies .

• Functional Consequences: The editing-derived amino acid changes likely affect protein structure, stability, interactions, and ultimately function, potentially influencing BLCAP's tumor-suppressive activities.

• Regulatory Network Insights: The BLCAP transcript serves as a model system for studying the activity and regulation of ADAR (Adenosine Deaminase Acting on RNA) enzymes, which catalyze A-to-I editing events. Both ADAR1 and ADAR2 cooperatively edit this transcript, with some site preferences observed (e.g., K/R site is preferentially edited by ADAR2) .

The study of BLCAP editing provides a valuable window into both the specific functions of this important tumor suppressor and the broader mechanisms of RNA editing regulation in normal physiology and cancer.

How can researchers experimentally detect and quantify RNA editing sites in BLCAP transcripts?

To detect and quantify RNA editing sites in BLCAP transcripts, researchers should employ a systematic methodological approach:

• RT-PCR and Sanger Sequencing:

  • Extract total RNA from tissues or cell lines

  • Synthesize cDNA using reverse transcriptase

  • Amplify BLCAP transcripts using specific primers flanking known or potential editing sites

  • Perform direct Sanger sequencing of the PCR products

  • Identify editing sites by mixed A/G peaks in chromatograms (since inosine is read as guanosine during sequencing)

  • Estimate editing levels by peak height ratios

• Cloning and Colony Sequencing:

  • Clone the RT-PCR products into appropriate vectors

  • Transform bacteria and pick multiple colonies (minimum 30-50 per sample)

  • Sequence individual clones

  • Calculate editing frequency as the percentage of clones showing edited sequences

• Next-Generation Sequencing (NGS):

  • Design RNA-seq experiments with sufficient depth to detect low-frequency editing events

  • Use strand-specific library preparation methods

  • Analyze data with specialized computational pipelines designed to identify RNA editing sites

  • Apply statistical filters to distinguish true editing events from sequencing errors

• Site-Specific Quantitative Methods:

  • Design SNP genotyping assays or qPCR assays that discriminate between edited and unedited sequences

  • Use restriction enzymes that differentially cleave edited versus unedited sequences

  • Implement droplet digital PCR (ddPCR) for absolute quantification of editing ratios

• Control Measures:

  • Always sequence genomic DNA to confirm that observed changes represent true editing events rather than genetic polymorphisms

  • Include known edited and unedited controls

  • Validate findings using multiple technical approaches

By combining these methods, researchers can comprehensively characterize the editing landscape of BLCAP transcripts across different tissues, disease states, and experimental conditions, as demonstrated in studies comparing editing levels between normal and cancerous tissues .

What is the relationship between ADAR enzymes and BLCAP RNA editing patterns?

The relationship between ADAR (Adenosine Deaminase Acting on RNA) enzymes and BLCAP RNA editing patterns is complex and highly regulated:

• Cooperative Editing by Multiple ADARs:

  • Both ADAR1 and ADAR2 contribute to editing the BLCAP transcript

  • The specific editing sites show different preferences for ADAR enzymes:

    • Y/C site: Edited by both ADAR1 and ADAR2, with a major contribution from ADAR1

    • K/R site: Preferentially edited by ADAR2

• Experimental Verification of ADAR Specificity:

  • Overexpression studies in HEK 293T cells demonstrate that both ADAR1 and ADAR2 can edit BLCAP transcripts, but with different efficiencies at various sites

  • The editing patterns observed correlate with the expression levels of specific ADAR enzymes

• Tissue-Specific Editing Patterns:

  • Different tissues show varying levels of BLCAP editing, which likely reflects:

    • Differential expression of ADAR1 and ADAR2 across tissues

    • Tissue-specific regulatory factors that modulate ADAR activity

    • Structural accessibility of the BLCAP transcript in different cellular contexts

• Dysregulation in Cancer:

  • Reduced BLCAP editing observed in cancerous tissues may result from:

    • Altered expression of ADARs in tumors

    • Changes in the regulatory network of proteins and RNAs that modulate ADAR function

    • Structural alterations in BLCAP transcripts affecting ADAR accessibility

• Methodological Approaches to Study ADAR-BLCAP Interactions:

  • ADAR knockdown/knockout experiments to assess specific contributions

  • ADAR overexpression studies to enhance editing at specific sites

  • RNA structure analysis to identify double-stranded regions required for ADAR activity

  • RNA-protein interaction assays to directly measure ADAR binding to BLCAP transcripts

This complex relationship between ADARs and BLCAP editing provides a valuable model system for studying RNA editing regulation more broadly, with implications for understanding how these processes are dysregulated in cancer .

How does BLCAP expression correlate with tumor progression and patient prognosis?

BLCAP expression demonstrates complex associations with tumor progression and patient outcomes across multiple dimensions:

• Expression Level Dynamics:

  • Loss of BLCAP mRNA expression correlates with invasive potential in urothelial carcinomas (UCs)

  • BLCAP protein expression is generally lost with tumor progression

  • Paradoxically, BLCAP is overexpressed in approximately 20% of UC cases examined

• Prognostic Significance:

  • Overexpression of BLCAP is linked with poor survival in bladder cancer patients

  • This suggests BLCAP may have prognostic value as a biomarker in certain cancer contexts

• Correlation with Malignancy Grade:

  • A correlation exists between decreased BLCAP editing levels (particularly at Q/R and K/R sites) and increased histological grade of malignancy in pediatric astrocytomas

  • This suggests editing status may provide additional prognostic information beyond mere expression levels

• Differential Cancer Type Patterns:

  • Downregulation of BLCAP expression has been observed in multiple cancer types including:

    • Bladder cancer

    • Renal cancer

    • Cervical cancer

    • Human tongue carcinoma

    • Osteosarcoma

• RNA Editing and Tumor Progression:

  • A general decrease in BLCAP editing is observed in astrocytomas, bladder cancer, and colorectal cancer compared to normal tissues

  • The editing events may generate protein isoforms with different functional properties, potentially influencing tumor behavior

• Functional Implications:

  • Experimental overexpression of BLCAP in cancer cell lines (HeLa and Tca8113) inhibits cell growth and induces apoptosis

  • This suggests the downregulation observed in tumors may contribute to cancer progression by removing growth inhibitory effects

These multifaceted relationships highlight BLCAP's potential value as both a prognostic biomarker and a therapeutic target, with changes in both expression level and editing status providing insight into tumor behavior and patient outcomes.

What molecular mechanisms underlie BLCAP's tumor suppressive functions?

The molecular mechanisms through which BLCAP exerts its tumor suppressive functions involve multiple cellular pathways and processes:

• Cell Cycle Regulation:

  • BLCAP appears to influence cell cycle progression, potentially through interactions with cyclins or cyclin-dependent kinases

  • Overexpression studies in cancer cell lines demonstrate inhibition of cell growth, suggesting BLCAP may induce cell cycle arrest

• Apoptosis Induction:

  • BLCAP overexpression in cervical cancer HeLa cells and tongue carcinoma Tca8113 cell lines induces apoptosis

  • This pro-apoptotic activity may involve activation of intrinsic or extrinsic apoptotic pathways

  • Potential mechanisms include regulation of Bcl-2 family proteins or activation of caspase cascades

• Protein-Protein Interactions:

  • Computer-based searches have identified potential functional domains in BLCAP

  • These domains may mediate interactions with other cellular proteins involved in growth control, apoptosis, or signal transduction

• RNA Editing Effects:

  • A-to-I editing of BLCAP transcripts creates protein isoforms with altered amino acid sequences

  • Specific editing events (Y/C, Q/R, K/R) may modify protein function or interactions

  • Different edited isoforms may have distinct tumor suppressive capacities

• Cellular Localization:

  • BLCAP protein expression and cellular localization patterns differ between benign bladder urothelium and urothelial carcinomas

  • These localization differences may impact BLCAP's interactions with other cellular components

• Gene Expression Regulation:

  • BLCAP may influence the expression of other genes involved in cell proliferation, survival, or motility

  • This could occur through direct or indirect effects on transcription factors or signaling pathways

Understanding these mechanisms provides potential avenues for therapeutic intervention, either by restoring BLCAP function in cancers where it is downregulated or by targeting downstream pathways in tumors that have lost BLCAP expression.

How can BLCAP be utilized as a biomarker in cancer diagnostics and prognosis?

BLCAP offers multiple properties that make it valuable as a cancer biomarker, with specific methodological approaches for implementation:

• Expression Level Assessment:

  • Develop standardized immunohistochemistry (IHC) protocols for BLCAP detection in tissue samples

  • Establish scoring systems to categorize expression as negative, low, moderate, or high

  • Correlate expression levels with clinicopathological parameters and patient outcomes

  • BLCAP protein expression is lost with tumor progression in some contexts, while approximately 20% of cases show overexpression linked to poor survival

• RNA Editing Status Analysis:

  • Implement RT-PCR followed by direct sequencing or cloning-based approaches to assess editing at key sites (Y/C, Q/R, K/R)

  • Correlate editing levels with tumor grade and patient outcomes

  • Develop quantitative assays (qPCR or digital PCR) for specific edited isoforms

  • Decreased editing levels correlate with increasing histological grade of malignancy in pediatric astrocytomas

• Multi-Parameter Biomarker Panels:

  • Combine BLCAP expression/editing assessment with other established biomarkers

  • Develop integrated scoring systems that incorporate multiple parameters

  • Use machine learning approaches to identify optimal biomarker combinations

• Liquid Biopsy Applications:

  • Investigate BLCAP mRNA or protein detection in circulation (blood, urine) for non-invasive diagnostics

  • Develop sensitive assays for detecting tumor-specific BLCAP isoforms in body fluids

  • Monitor changes in BLCAP expression/editing during treatment as a potential response marker

• Clinical Implementation Strategy:

  • Validate in large, prospective clinical cohorts with long-term follow-up

  • Standardize testing methodologies across laboratories

  • Establish clear cut-off values for clinical decision-making

  • Develop quality control measures to ensure reproducible results

This comprehensive approach to BLCAP biomarker development leverages both expression level data and editing status information to maximize diagnostic and prognostic utility across different cancer types. The established correlation between BLCAP alterations and tumor characteristics provides a strong foundation for its clinical application as a biomarker .

How can researchers design experiments to investigate the functional consequences of specific RNA editing events in BLCAP?

To elucidate the functional consequences of specific RNA editing events in BLCAP, researchers should implement a comprehensive experimental strategy:

• Site-Directed Mutagenesis Approach:

  • Generate expression constructs that mimic edited and unedited BLCAP variants:

    • Wild-type (unedited) BLCAP

    • Y/C edited variant (Tyrosine to Cysteine)

    • Q/R edited variant (Glutamine to Arginine)

    • K/R edited variant (Lysine to Arginine)

    • Combinations of multiple editing events

  • Express these variants in appropriate cell models and assess:

    • Protein stability and half-life

    • Subcellular localization

    • Effects on cell proliferation, migration, and apoptosis

    • Differential protein-protein interactions

• CRISPR-Based Editing Manipulation:

  • Use CRISPR-Cas13 RNA editing systems to specifically modify BLCAP transcripts at editing sites

  • Develop CRISPR strategies to alter ADAR binding sites in the genomic BLCAP locus

  • Create cell lines with RNA editing-resistant BLCAP variants

• Structural Biology Investigations:

  • Perform structural analyses (NMR, X-ray crystallography) of edited and unedited BLCAP proteins

  • Use computational modeling to predict structural changes resulting from amino acid substitutions

  • Identify potential functional domains affected by editing

• Transcriptome and Proteome Analysis:

  • Conduct RNA-seq and proteomics on cells expressing different BLCAP variants

  • Identify downstream pathways differentially affected by edited versus unedited BLCAP

  • Perform pathway enrichment analysis to contextualize functional consequences

• In Vivo Models:

  • Generate transgenic mouse models expressing edited or unedited BLCAP variants

  • Examine tissue-specific phenotypes

  • Investigate susceptibility to cancer development

  • Evaluate response to carcinogens or cancer therapies

• Tissue-Specific Context:

  • Analyze editing patterns in different tissues and correlate with BLCAP function

  • Investigate whether tissue-specific factors influence the functional outcomes of BLCAP editing

  • Consider how the varying editing levels observed across tissues (e.g., 27.6% at Y/C site in bladder versus 5.1% in heart) might reflect tissue-specific requirements for BLCAP function

This multifaceted approach will provide comprehensive insights into how RNA editing modulates BLCAP function and its implications for tumor suppression in different cellular contexts.

What are the challenges and potential solutions in developing therapeutic approaches targeting BLCAP in cancer?

Developing therapeutic approaches targeting BLCAP in cancer presents several challenges along with potential solutions:

• Dual Role Complexity:

  • Challenge: BLCAP displays a paradoxical behavior—generally downregulated in tumors but overexpressed in ~20% of cases where it correlates with poor prognosis .

  • Solution: Implement precision medicine approaches with comprehensive biomarker testing to identify which patients would benefit from BLCAP restoration versus inhibition.

• RNA Editing Modulation:

  • Challenge: Targeting specific BLCAP editing events therapeutically requires precise manipulation of ADAR activity, which affects numerous transcripts.

  • Solution: Develop site-specific RNA editing tools using modified CRISPR-Cas13 systems or antisense oligonucleotides that can shield specific editing sites from ADAR enzymes.

• Delivery Systems:

  • Challenge: Efficiently delivering BLCAP-modulating agents to target tissues.

  • Solution: Explore nanoparticle formulations, exosome-based delivery, or targeted viral vectors to achieve tissue-specific BLCAP modulation.

• Restoration of Expression:

  • Challenge: Effectively restoring BLCAP expression in tumors where it is downregulated.

  • Solution:

    • Investigate epigenetic modifiers that might reverse silencing of the BLCAP gene

    • Develop mRNA therapeutics containing the BLCAP coding sequence

    • Design viral vectors for BLCAP gene therapy approaches

• Functional Pathway Targeting:

  • Challenge: Direct targeting of BLCAP may be difficult due to its small size and limited structural information.

  • Solution: Identify and target downstream effectors of BLCAP's tumor suppressive function or synthetic lethal partners in BLCAP-deficient tumors.

• Monitoring Treatment Response:

  • Challenge: Assessing the therapeutic efficacy of BLCAP-targeting approaches.

  • Solution: Develop companion diagnostics that can monitor BLCAP expression, editing status, or downstream pathway activation in liquid biopsies or serial tumor samples.

• Resistance Mechanisms:

  • Challenge: Tumors may develop resistance to BLCAP-based therapies.

  • Solution: Investigate combination therapies targeting multiple pathways and identify potential resistance mechanisms in preclinical models.

• Clinical Trial Design:

  • Challenge: Designing appropriate clinical trials for BLCAP-targeted therapies.

  • Solution: Implement basket or umbrella trial designs that group patients based on BLCAP status rather than traditional cancer type classifications.

These strategies address the multifaceted challenges of BLCAP-targeted therapy development, leveraging our understanding of its complex biology while acknowledging the practical hurdles in therapeutic implementation.

How can comparative studies of BLCAP across different species contribute to understanding its evolutionary significance and functional conservation?

Comparative studies of BLCAP across species provide valuable insights into its evolutionary significance and functional conservation, with several key methodological approaches:

• Sequence Conservation Analysis:

  • Perform multiple sequence alignments of BLCAP proteins from diverse species

  • Calculate evolutionary conservation scores for each amino acid position

  • Identify highly conserved domains that likely mediate critical functions

  • Pay particular attention to conservation around known RNA editing sites (Y/C, Q/R, K/R)

  • The high conservation of BLCAP across species (as evidenced by availability of recombinant proteins from diverse organisms including Didelphis marsupialis virginiana) suggests important functional roles

• RNA Editing Conservation:

  • Compare RNA editing patterns of BLCAP across different species

  • Determine whether editing sites are positionally conserved

  • Assess whether editing frequencies show species-specific patterns

  • Investigate the evolution of editing-dependent regulatory mechanisms

• Expression Pattern Comparison:

  • Analyze tissue-specific expression profiles of BLCAP across species

  • Compare developmental expression patterns

  • Evaluate whether expression changes in disease states (particularly cancer) are conserved

  • Identify conserved regulatory elements in BLCAP promoter regions

• Functional Studies in Model Organisms:

  • Generate BLCAP knockout/knockin models in evolutionary distinct species (e.g., mouse, zebrafish, Drosophila)

  • Compare phenotypes to identify conserved versus species-specific functions

  • Perform cross-species rescue experiments (e.g., can human BLCAP rescue function in zebrafish knockouts?)

  • Test whether editing-site mutations produce similar phenotypes across species

• Protein Interaction Network Evolution:

  • Identify BLCAP-interacting proteins in different species using techniques like:

    • Yeast two-hybrid screening

    • Co-immunoprecipitation followed by mass spectrometry

    • Proximity labeling approaches (BioID, APEX)

  • Compare interaction networks to identify conserved and divergent interacting partners

  • Assess whether editing-dependent interactions are evolutionarily conserved

• Cancer-Related Functions Across Species:

  • Compare BLCAP expression and editing patterns in naturally occurring or induced cancers across species

  • Determine whether tumor suppressive functions are conserved

  • Investigate species-specific differences in BLCAP's role in cancer development

This multi-layered comparative approach provides insights into BLCAP's fundamental biological importance, the selective pressures that have shaped its evolution, and how this evolutionary context informs our understanding of its role in human cancer. The high conservation of this protein, from marsupials to humans, strongly suggests it performs critical cellular functions that have been maintained throughout mammalian evolution .

Table: BLCAP RNA Editing Levels in Different Human Tissues

Data compiled from research findings showing tissue-specific editing patterns of BLCAP transcripts .

Table: Comparison of BLCAP Editing in Normal vs. Cancer Tissues

*Correlation observed between decreased editing levels and increased histological grade of malignancy in pediatric astrocytomas.
**With some exceptions in specific cancer cell lines.
Data summarized from research findings comparing editing levels between normal and cancerous tissues .

Table: Properties of Recombinant Didelphis Marsupialis Virginiana BLCAP Protein

Data from product specifications for recombinant BLCAP protein .