Recombinant Human E3 ubiquitin-protein ligase TRIM13 (TRIM13)

What is the basic structure and localization of TRIM13?

TRIM13 is a transmembrane E3 ubiquitin ligase that contains three conserved domains in its N-terminus: RING (R), B-Box (B), and coiled-coil (CC) domains, with a transmembrane domain (TM) in its C-terminus . This structure is characteristic of TRIM family proteins, though the presence of a transmembrane domain is distinctive for TRIM13. Subcellular localization studies demonstrate that TRIM13 is predominantly an ER-resident protein, with specific localization to the ER membrane . This localization is critical for its function, as the transmembrane domain allows TRIM13 to interact with other membrane-bound proteins including STING, a key mediator of innate immune responses to cytosolic DNA .

For researchers studying TRIM13, it's essential to consider that the protein's functionality depends on its correct localization, and experimental designs should account for this when expressing recombinant constructs. Using confocal microscopy with appropriate ER markers (such as calnexin or PDI) can confirm proper localization of wild-type or mutant TRIM13 proteins in experimental systems.

How is TRIM13 expression regulated in normal versus cancerous tissues?

TRIM13 expression shows significant variation between normal and cancerous tissues. Multiple studies have demonstrated that TRIM13 is downregulated in various human malignancies. In lung cancer specifically, TRIM13 exhibits significantly reduced expression in tumor tissues compared to adjacent normal tissues, as confirmed by both qRT-PCR and immunohistochemistry analyses . Analysis of 574 lung adenocarcinoma (LUAD) samples from the TCGA database showed consistently lower TRIM13 mRNA expression in tumor samples .

The genomic basis for this downregulation relates to TRIM13's location on chromosome 13q14, a region frequently deleted in various malignancies . This deletion pattern has been observed in esophageal cancer, ovarian cancer, leukemia, and breast cancer, suggesting that TRIM13 downregulation may be a common feature across multiple cancer types .

What are the established cellular functions of TRIM13?

TRIM13 exhibits several critical cellular functions that have been experimentally validated:

• Autophagy Induction: TRIM13 has been shown to induce autophagy when ectopically expressed in cells. Domain mapping studies have demonstrated that the coiled-coil (CC) domain is essential for this autophagy-inducing function .

• ER Stress Regulation: TRIM13 is stabilized during ER stress and plays a regulatory role in the initiation of autophagy under these conditions. It interacts with p62/SQSTM1 and co-localizes with DFCP1, suggesting involvement in autophagosome formation .

• Tumor Suppression: Functional studies have established TRIM13 as a tumor suppressor. Overexpression of TRIM13 in lung cancer cell lines (A549 and H157) significantly inhibits cell proliferation, migration, and invasion, as demonstrated through CCK-8 and transwell assays .

• Ubiquitination and Protein Degradation: TRIM13 functions as an E3 ubiquitin ligase that targets specific proteins for ubiquitination and subsequent degradation. For example, it interacts with RPS27A, increasing its ubiquitination and degradation .

• Immune Response Regulation: TRIM13 regulates innate immune responses to DNA viruses by mediating the ubiquitination of STING, a key signaling protein in the cytosolic DNA sensing pathway .

Researchers studying TRIM13 functions should design experiments with appropriate controls to distinguish between these diverse functions, particularly when investigating a novel cellular context or interacting partner.

How does TRIM13 regulate the STING-mediated DNA sensing pathway in innate immunity?

TRIM13 plays a sophisticated regulatory role in the STING-mediated DNA sensing pathway through multiple mechanisms. Research has demonstrated that TRIM13 interacts directly with STING (stimulator of interferon genes) through their respective transmembrane domains . This interaction occurs primarily at the ER membrane under resting conditions, with colocalization observed in both RAW264.7 and HeLa cell lines .

Upon DNA virus infection (such as with HSV-1), this colocalization is attenuated, suggesting a dynamic regulatory relationship dependent on infection status . Mechanistically, TRIM13 mediates K6-linked polyubiquitination of STING, which has two major consequences: 1) it decelerates STING's exit from the ER, and 2) it accelerates ER-initiated degradation of STING .

In TRIM13-deficient cells, researchers have observed accelerated removal of STING from the ER after HSV-1 or VACV infection . This finding was confirmed in both TRIM13-deficient BMDMs (bone marrow-derived macrophages) and MEFs (mouse embryonic fibroblasts) . The consequence of this accelerated ER exit is enhanced STING-dependent innate immune signaling and increased inflammatory cytokine production.

For researchers investigating this pathway, it's important to note that TRIM13's effect on STING cannot be fully reversed by proteasome inhibitors like MG132, suggesting that TRIM13 may also affect STING degradation through alternative pathways such as ER-phagy . Additionally, the specificity of TRIM13's ubiquitination of STING should be considered in the context of other E3 ligases known to target STING, including RNF5, TRIM29, TRIM30a, TRIM56, MUL1, TRIM32, and RNF26, which mediate different types of ubiquitin linkages (K48, K63, K11) at various lysine residues on STING .

What is the molecular mechanism by which TRIM13 suppresses lung cancer progression?

TRIM13 suppresses lung cancer progression through a complex molecular mechanism centered on its interaction with RPS27A and subsequent regulation of the NF-κB signaling pathway. Researchers have elucidated this mechanism through a series of biochemical and functional studies:

• TRIM13-RPS27A Interaction: TRIM13 directly binds to RPS27A (Ribosomal Protein S27A), as confirmed by co-immunoprecipitation (Co-IP) assays in lung cancer cells .

• Ubiquitination and Degradation of RPS27A: Following binding, TRIM13 promotes the ubiquitination of RPS27A, leading to its proteasomal degradation. This was demonstrated by experiments showing that TRIM13 overexpression results in a marked increase in ubiquitin co-immunoprecipitated with RPS27A .

• Impact on Protein Stability: TRIM13 overexpression enhances RPS27A downregulation induced by the protein synthesis inhibitor cycloheximide (CHX) in lung cancer cells, confirming that TRIM13 affects RPS27A protein stability .

• NF-κB Pathway Inhibition: The degradation of RPS27A leads to inhibition of the NF-κB signaling pathway, which is a key promoter of cancer cell proliferation and metastasis . This inhibitory effect was demonstrated to be reversible when RPS27A was overexpressed alongside TRIM13 .

• Functional Consequences: Through these molecular interactions, TRIM13 overexpression significantly inhibits lung cancer cell proliferation, migration, and invasion in vitro, and suppresses tumor growth in xenograft mouse models in vivo .

For researchers studying this mechanism, it's important to investigate the specificity of this interaction by employing appropriate controls and examining potential effects on other signaling pathways. Understanding the structural determinants of the TRIM13-RPS27A interaction could also provide insights for developing targeted therapeutic approaches.

What is the role of TRIM13 in age-related autoinflammation and autoimmunity?

TRIM13 deficiency has been linked to age-related autoinflammation in mouse models, providing important insights into potential roles in human autoimmune and autoinflammatory conditions. Research has shown that while young TRIM13-deficient mice (5 months old) do not display significant inflammatory phenotypes, older knockout mice (10 months old) develop extensive inflammatory cell infiltrations in multiple organs .

Specifically, histological examination of 10-month-old TRIM13-deficient mice revealed:

• Inflammatory cell infiltration in the lungs (around bronchia or vessels) in 87.5% of examined mice

• Kidney inflammation in 75.0% of mice

• Liver inflammation in a significant proportion of mice

• Milder inflammatory cell infiltration in the heart and brain

This age-dependent autoinflammation in TRIM13-deficient mice parallels observations with mutations in other STING-interacting proteins (STIM1, TMEM203, and C9ORF72) that have been implicated in autoinflammatory and autoimmune diseases in humans .

The molecular basis for this phenotype appears to be connected to TRIM13's regulatory role in the STING pathway, which controls innate immune responses to cytosolic DNA. Without TRIM13's restraining influence on STING activation, chronic or aberrant activation of DNA-sensing pathways may occur, leading to excessive inflammatory cytokine production over time .

For researchers investigating these connections, it's valuable to consider:

• Age as a critical variable in study design when using TRIM13-deficient models

• Examination of multiple organs for inflammatory phenotypes

• Correlation with STING activation markers and downstream cytokine production

• Potential therapeutic interventions targeting the STING pathway

What are the best experimental approaches to study TRIM13's E3 ligase activity?

Studying TRIM13's E3 ligase activity requires specialized techniques to capture the dynamic process of protein ubiquitination. Based on established research protocols, the following methodological approaches are recommended:

• In vitro Ubiquitination Assays: These assays require purified recombinant TRIM13 (focusing on the RING domain), E1 and E2 enzymes, ubiquitin, ATP, and the substrate protein of interest (e.g., RPS27A or STING). The reaction products can be analyzed by SDS-PAGE followed by Western blotting using anti-ubiquitin antibodies. This approach allows researchers to determine whether TRIM13 directly catalyzes ubiquitin transfer and identify the type of ubiquitin linkage formed .

• Cellular Ubiquitination Assays: These involve co-expressing TRIM13 with the substrate protein and HA-tagged or FLAG-tagged ubiquitin in cells, followed by immunoprecipitation of the substrate and detection of ubiquitinated forms by Western blotting. This approach was successfully used to demonstrate TRIM13-mediated ubiquitination of RPS27A in lung cancer cells .

• Ubiquitin Chain Linkage Analysis: To determine the specific type of ubiquitin chain linkage (K6, K11, K48, K63, etc.) mediated by TRIM13, researchers can use ubiquitin mutants with specific lysine-to-arginine substitutions or linkage-specific antibodies. For TRIM13, K6-linked polyubiquitination of STING has been demonstrated .

• Cycloheximide Chase Assays: To assess the impact of TRIM13-mediated ubiquitination on protein stability, researchers can perform cycloheximide chase assays, treating cells with the protein synthesis inhibitor cycloheximide and monitoring the degradation rate of the substrate protein in the presence or absence of TRIM13 .

• Proteasome Inhibition Experiments: Using proteasome inhibitors like MG132 can help determine whether TRIM13-mediated ubiquitination leads to proteasomal degradation of the substrate. If protein levels are not fully restored by MG132, alternative degradation pathways (such as autophagy) should be investigated .

• TRIM13 Domain Mutants: Generating TRIM13 constructs with mutations in the RING domain (which typically abolishes E3 ligase activity) serves as an important negative control to confirm that observed ubiquitination is directly dependent on TRIM13's catalytic activity .

When designing these experiments, researchers should consider using appropriate tags that do not interfere with TRIM13's membrane localization or E3 ligase activity.

How can researchers effectively overexpress or knock down TRIM13 in experimental systems?

Effective manipulation of TRIM13 expression levels is crucial for studying its functions. Based on successful approaches documented in the literature, researchers should consider the following methods:

For TRIM13 Overexpression:

• Plasmid Selection: The choice of expression vector should be compatible with the experimental system. For TRIM13, vectors with CMV or EF1α promoters have been effectively used for strong expression in mammalian cells . Consider using inducible systems (e.g., Tet-On) for temporal control of expression.

• Cell Transfection Methods: For transient transfection of TRIM13 in lung cancer cell lines (A549 and H157), lipid-based transfection reagents have been successfully employed . The transfection efficiency should be verified by qRT-PCR and Western blotting.

• Viral Delivery Systems: For cell types that are difficult to transfect or for in vivo studies, lentiviral or adenoviral delivery of TRIM13 expression constructs provides higher efficiency. This approach has been used effectively in xenograft mouse models of lung cancer .

• Considerations for TRIM13 Constructs:

  • Include appropriate epitope tags (e.g., FLAG, HA, or Myc) that do not interfere with TRIM13's transmembrane domain

  • Consider including domain deletion mutants (ΔRING, ΔTM) as controls to study domain-specific functions

  • Use codon-optimized sequences for enhanced expression

For TRIM13 Knockdown/Knockout:

• siRNA/shRNA Approaches: Small interfering RNA (siRNA) can be used for transient knockdown, while short hairpin RNA (shRNA) delivered via lentiviral vectors provides stable knockdown. Multiple siRNA sequences targeting different regions of TRIM13 mRNA should be tested to identify the most efficient knockdown .

• CRISPR-Cas9 Gene Editing: For complete knockout of TRIM13, CRISPR-Cas9 technology has been successfully employed to generate TRIM13-deficient cell lines and mouse models . Multiple guide RNAs should be designed and validated for efficient targeting.

• Verification Methods:

  • Confirm knockdown/knockout efficiency at both mRNA level (qRT-PCR) and protein level (Western blotting)

  • Examine potential off-target effects by rescue experiments with TRIM13 constructs resistant to the siRNA/shRNA or by using multiple independent CRISPR clones

• Considerations for In Vivo Models: When working with TRIM13 knockout mice, researchers should be aware of the age-dependent phenotypes. Young TRIM13-deficient mice (5 months) may not display obvious phenotypes, while older mice (10 months) develop autoinflammatory conditions .

For all expression manipulation approaches, appropriate controls (empty vector, non-targeting siRNA/shRNA, or wild-type cells/animals) should be included in experimental designs.

What are the recommended methods for studying TRIM13's impact on autophagy and ER stress?

Investigating TRIM13's role in autophagy and ER stress requires specialized experimental approaches that capture these dynamic cellular processes. Based on established protocols, the following methods are recommended:

For Studying TRIM13's Role in Autophagy:

• Autophagosome Visualization:

  • Fluorescence microscopy using GFP-LC3 to monitor autophagosome formation in cells expressing wild-type or mutant TRIM13

  • Transmission electron microscopy (TEM) to directly visualize autophagic structures at ultrastructural level

  • Co-localization studies of TRIM13 with autophagy markers such as DFCP1 and p62/SQSTM1

• Autophagic Flux Assessment:

  • Western blotting for LC3-I to LC3-II conversion in the presence/absence of lysosomal inhibitors (e.g., bafilomycin A1, chloroquine)

  • Monitoring degradation of autophagy substrates such as p62/SQSTM1

  • Tandem fluorescent-tagged LC3 (mRFP-GFP-LC3) assay to distinguish between autophagosomes and autolysosomes

• Domain Requirement Analysis:

  • Transfection with TRIM13 constructs lacking specific domains (particularly the coiled-coil domain that has been shown to be required for autophagy induction)

  • Quantification of autophagic markers to determine which domains are essential for TRIM13's pro-autophagy function

For Studying TRIM13's Role in ER Stress:

• ER Stress Induction:

  • Treatment with established ER stress inducers (tunicamycin, thapsigargin, DTT, or glucose deprivation)

  • Monitoring TRIM13 protein stabilization during ER stress by Western blotting

• ER Stress Markers Assessment:

  • qRT-PCR and Western blotting for canonical ER stress markers (BiP/GRP78, CHOP, XBP1 splicing, phosphorylation of eIF2α and PERK)

  • Analysis of these markers in cells with TRIM13 overexpression or knockdown/knockout

• ER-Associated Degradation (ERAD) Analysis:

  • Pulse-chase experiments to monitor degradation of ERAD substrates in the presence/absence of TRIM13

  • Co-immunoprecipitation to identify TRIM13 interactions with ERAD machinery components

• ER Morphology Examination:

  • Immunofluorescence microscopy using ER markers (calnexin, PDI, Sec61)

  • ER-tracker dyes combined with live-cell imaging

  • TEM to analyze changes in ER structure in TRIM13-deficient cells

• Functional Consequence Assessment:

  • Clonogenic assays to determine the impact of TRIM13-mediated ER stress responses on cell survival

  • Analysis of ER stress-induced cell death pathways (apoptosis vs. non-apoptotic cell death)

When designing these experiments, researchers should consider using appropriate cell types where TRIM13 is endogenously expressed at detectable levels. Additionally, time-course experiments are crucial for capturing the dynamic nature of both autophagy and ER stress responses.

How does TRIM13 expression correlate with cancer prognosis and potential therapeutic strategies?

TRIM13 expression has been demonstrated to have significant prognostic value in multiple cancer types, with important implications for therapeutic development. Based on current research findings:

Prognostic Correlations:

Therapeutic Implications:

• TRIM13/RPS27A/NF-κB Axis: Targeting this signaling axis represents a promising therapeutic strategy for lung cancer treatment. The mechanism involves TRIM13's regulation of RPS27A degradation, which in turn inhibits the NF-κB pathway . Therapeutic approaches could focus on:

  • Enhancing TRIM13 expression or activity

  • Directly targeting RPS27A

  • Inhibiting NF-κB pathway components downstream of this interaction

• Multiple Myeloma Approach: Research has shown that TRIM13 downregulation in multiple myeloma (MM) reduces cell cycle progression and proliferation by inhibiting NF-κB pathway activity and the 20S proteasome . This suggests TRIM13 as a potential therapeutic target specifically for MM treatment.

• Combination Therapy Potential: The involvement of TRIM13 in both proteasomal and autophagic degradation pathways suggests that combining TRIM13-targeting approaches with proteasome inhibitors or autophagy modulators might yield synergistic therapeutic effects.

• Gene Therapy Considerations: Given TRIM13's location on chromosome 13q14, a region frequently deleted in various malignancies, gene therapy approaches to restore TRIM13 expression in tumors with 13q14 deletions could be explored.

For researchers and clinicians exploring these therapeutic avenues, it's important to consider cancer-specific contexts, as TRIM13's role appears to vary between cancer types. Additionally, patient stratification based on TRIM13 expression levels may be valuable for personalizing therapeutic approaches.

What is the potential for targeting TRIM13 in inflammatory and autoimmune disorders?

Research on TRIM13's role in regulating innate immune responses suggests significant potential for therapeutic targeting in inflammatory and autoimmune conditions. The evidence indicates:

• Age-Related Autoinflammation in TRIM13 Deficiency: TRIM13-deficient mice develop age-dependent inflammatory phenotypes affecting multiple organs, including the lung, liver, kidney, heart, and brain . This suggests that dysregulation of TRIM13 may contribute to inflammaging and autoimmune conditions in humans.

• Mechanistic Link to STING Pathway: TRIM13 regulates the STING-mediated DNA sensing pathway by controlling STING's ER exit and degradation through K6-linked polyubiquitination . This pathway is increasingly recognized as central to various autoimmune and autoinflammatory diseases.

• Connection to Known Autoimmune Conditions: The phenotypes observed in TRIM13-deficient mice parallel those seen with mutations in other STING-interacting proteins (STIM1, TMEM203, and C9ORF72) that have been implicated in human autoinflammatory and autoimmune diseases .

Therapeutic Targeting Strategies:

• TRIM13 Enhancement: For conditions involving hyperactive STING signaling, approaches to enhance TRIM13 expression or activity could potentially dampen excessive inflammatory responses. This might include:

  • Small molecule activators of TRIM13's E3 ligase activity

  • Gene therapy approaches to increase TRIM13 expression

  • Peptide mimetics that replicate TRIM13's interaction with STING

• Targeting the TRIM13-STING Interaction: Developing compounds that modulate the specific interaction between TRIM13 and STING could provide precise control over this signaling pathway. Structural studies of the transmembrane domains that mediate this interaction would be valuable for drug design.

• Age-Dependent Intervention: Given the age-dependent nature of autoinflammation in TRIM13-deficient models, preventive interventions might be most effective when applied before the onset of inflammatory manifestations.

• Biomarker Development: TRIM13 expression or activity levels could potentially serve as biomarkers for stratifying patients with inflammatory or autoimmune conditions, identifying those who might benefit from therapies targeting this pathway.

For researchers in this field, it's important to consider that therapeutic approaches would need to achieve precise modulation rather than complete inhibition or activation of TRIM13, as both excessive and insufficient activity appear to have pathological consequences.

What are the emerging roles of TRIM13 beyond cancer and immunity?

While TRIM13's functions in cancer suppression and immune regulation have been well-documented, recent research has begun to uncover additional roles that expand our understanding of this versatile E3 ubiquitin ligase:

• ER-phagy Regulation: Emerging evidence suggests TRIM13 may participate in ER-phagy, a selective form of autophagy that degrades portions of the ER . This role extends beyond general autophagy induction and may represent a specialized function of TRIM13 in maintaining ER homeostasis.

• Proteasome Activity Modulation: Research in multiple myeloma has indicated that TRIM13 can regulate 20S proteasome activity , suggesting a broader role in protein quality control beyond its direct E3 ligase targets.

• Cross-talk with Other TRIM Family Members: Recent studies are beginning to reveal functional interactions and compensatory mechanisms between TRIM13 and other TRIM family proteins, indicating a complex network of regulation within this protein family.

• Neurological Function: The observation of mild inflammatory infiltration in the brains of aged TRIM13-deficient mice suggests potential roles in neurological function or neuroprotection that have yet to be fully explored.

Future research directions should include:

• Investigation of TRIM13's role in neurodegenerative diseases characterized by protein aggregation

• Exploration of potential functions in metabolic regulation, given its location in the ER

• Analysis of TRIM13 polymorphisms and their association with disease susceptibility

• Examination of post-translational modifications that regulate TRIM13 activity

What are the current technical challenges in TRIM13 research and how can they be addressed?

Researchers studying TRIM13 face several technical challenges that can limit progress in understanding its functions and developing therapeutic applications:

• Protein Detection Difficulties:

  • TRIM13 is often expressed at low levels, making endogenous protein detection challenging

  • The transmembrane domain can cause aggregation during sample preparation for Western blotting

  Solutions:

  • Use of more sensitive detection methods such as proximity ligation assays

  • Optimization of membrane protein extraction protocols

  • Development of higher-affinity antibodies specifically validated for TRIM13 detection

• Functional Analysis of Membrane-Associated E3 Ligases:

  • Standard in vitro ubiquitination assays may not fully recapitulate the membrane context required for TRIM13 function

  Solutions:

  • Development of membrane-based in vitro systems that preserve the lipid environment

  • Use of microsomal preparations instead of purified proteins for activity assays

  • Application of proximity-based labeling techniques (BioID, APEX) to identify substrates in their native context

• Complex Phenotypes in Animal Models:

  • Age-dependent phenotypes in TRIM13-deficient mice require long-term studies

  • Potential compensatory mechanisms from other TRIM family members

  Solutions:

  • Generation of conditional and tissue-specific knockout models

  • Development of acute TRIM13 inhibition methods for temporal control

  • Creation of double or triple knockout models with related TRIM proteins

• Translation to Human Disease:

  • Limited availability of patient samples with well-characterized TRIM13 status

  • Uncertainty about which model systems best recapitulate human TRIM13 biology

  Solutions:

  • Establishment of patient-derived xenografts or organoids

  • CRISPR-engineered human cell lines with patient-specific TRIM13 mutations

  • Integration of genetic, transcriptomic, and proteomic data from patient cohorts

Addressing these technical challenges will require interdisciplinary approaches combining structural biology, advanced imaging, proteomics, and genetic tools. Collaborative efforts between academic and industrial researchers may accelerate progress in overcoming these obstacles.

How can multi-omics approaches enhance our understanding of TRIM13 function and regulation?

Integrating multiple omics technologies offers powerful approaches to comprehensively understand TRIM13's functions and regulatory networks:

• Proteomics Applications:

  • Ubiquitinome Analysis: Quantitative proteomics using di-glycine remnant profiling can identify the complete set of proteins whose ubiquitination status changes upon TRIM13 manipulation, revealing direct and indirect substrates .

  • Interactome Mapping: Proximity-based labeling methods (BioID, APEX) combined with mass spectrometry can identify the TRIM13 protein interaction network within the ER membrane context.

  • Post-translational Modification Profiling: Analysis of how TRIM13 itself is regulated by phosphorylation, SUMOylation, or other modifications that might control its E3 ligase activity or stability.

• Transcriptomic Approaches:

  • RNA-Seq Analysis: Comparing transcriptional profiles of TRIM13-deficient versus wild-type cells under normal and stress conditions can reveal regulatory networks affected by TRIM13.

  • Single-Cell Transcriptomics: Examining cell-to-cell variability in responses to TRIM13 manipulation, particularly in immune or cancer cell populations.

  • Alternative Splicing Analysis: Investigating whether TRIM13 indirectly affects RNA processing through its regulation of stress responses.

• Genomic Strategies:

  • ChIP-Seq Following TRIM13 Manipulation: Identifying changes in transcription factor binding or epigenetic marks that result from TRIM13-mediated regulation of signaling pathways like NF-κB .

  • CRISPR Screens: Genome-wide or targeted CRISPR-Cas9 screens to identify genes that synthetically interact with TRIM13, revealing functional relationships and compensatory mechanisms.

  • SNP Association Studies: Analysis of TRIM13 genetic variants in large patient cohorts to identify associations with disease susceptibility or progression.

• Metabolomic Investigations:

  • Metabolic Profiling: Examining how TRIM13 manipulation affects cellular metabolism, particularly in cancer contexts where metabolic reprogramming is common.

  • Lipid Analysis: Given TRIM13's ER localization, investigating its potential impact on lipid composition and ER membrane properties.

• Integrative Multi-omics:

  • Network Analysis: Integration of protein, RNA, and metabolite data to construct comprehensive regulatory networks centered on TRIM13.

  • Systems Biology Modeling: Development of mathematical models to predict the dynamic behavior of TRIM13-regulated pathways under various conditions.

  • Temporal Multi-omics: Capturing time-course data across multiple molecular levels during cellular responses where TRIM13 plays a role.