ELOB Mouse
Description
Introduction to ELOB Mouse
ELOB Mouse refers to research models and recombinant proteins derived from the ELOB gene in mice, encoding elongin B, a regulatory subunit of the transcription factor B (SIII) complex. This complex facilitates RNA polymerase II transcription elongation by suppressing polymerase pausing . Studies in mice have elucidated its role in development, disease, and chemical interactions, with recombinant ELOB proteins used in biochemical assays .
Gene Structure and Function
The ELOB gene (also known as TCEB2) encodes a 141-amino acid protein (15.6 kDa) critical for transcriptional regulation. Key features include:
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Subunit role: Forms a heterodimer with elongin C, binding elongin A to enhance transcriptional activity .
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Regulatory interactions: Binds von Hippel-Lindau (VHL) tumor suppressor protein to inhibit transcription elongation .
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Alternative splicing: Produces isoforms with distinct tissue-specific expression .
Developmental and Cellular Functions
Tissue Expression Patterns
Chemical and Environmental Interactions
ELOB interacts with diverse compounds, affecting its expression or protein stability:
Modulators of ELOB Expression
Protein Stability Modulators
| Compound | Effect | Context | Source |
|---|---|---|---|
| Nutlin-3 + Dactinomycin | ↑ Secretion | Enhances ELOB protein release | |
| Ivermectin | ↓ Protein levels | Suppresses transcriptional activity |
Recombinant ELOB Proteins
Gene Knockout Models
| Model | Phenotype | Implication | Source |
|---|---|---|---|
| Elobl KO mice | Normal fecundity | Redundancy in ovarian development | |
| Nlrp2 KO mice | ↓ Pups/litter (5.4 ± 2.6) | Critical for fertility regulation |
Clinical and Pathological Relevance
ELOB is implicated in:
Product Specs
Transcription elongation factor B polypeptide 2, TCEB2, RNA polymerase IItranscription factor SIII subunit B, SIII p18, EloB, Elongin 18 kDa subunit.
MGSSHHHHHH SSGLVPRGSH MGSMDVFLMI RRHKTTIFTD AKESSTVFEL KRIVEGILKR PPEEQRLYKD DQLLDDGKTL GECGFTSQTA RPQAPATVGL AFRADDTFEA LRIEPFSSPP ELPDVMKPQD SGGSANEQAV Q
Q&A
Basic Research Questions
What experimental approaches are recommended for detecting ELOB expression in mouse tissues?
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Methodological guidance:
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Use Western blot with validated antibodies (e.g., Boster Bio A31718-1, reactive to mouse ELOB ) at 1:500–1:2000 dilutions. Include positive controls (e.g., mouse liver or kidney lysates) and validate via siRNA knockdown.
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For mRNA quantification, employ qRT-PCR with primers targeting Elob exons 2–4 (UCSC Genome Browser coordinates). Normalize to housekeeping genes (e.g., Gapdh, Actb).
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Reference multi-lab validation protocols from electrophysiology studies to ensure reproducibility, such as standardized tissue collection intervals post-euthanasia.
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How do I design a study to profile Elob expression across mouse developmental stages?
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Key considerations:
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Utilize RNA in situ hybridization with probes spanning Elob coding regions (NCBI Gene ID: 1914923 ).
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Stratify sampling by Theiler stages (TS13–TS28) and include at least three biological replicates per stage.
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Cross-reference expression data with public repositories (e.g., MGI’s Gene Expression Database ) to identify baseline patterns.
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Advanced Research Questions
How should researchers resolve contradictions in ELOB protein localization data across studies?
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Analytical framework:
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Perform subcellular fractionation followed by mass spectrometry to distinguish nuclear vs. cytoplasmic pools.
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Compare antibody specificity using knockout controls (e.g., Elob −/− mice) to rule out cross-reactivity .
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Apply statistical methods from multi-lab electrophysiology studies , such as permutation tests for inter-lab variability assessment.
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What strategies optimize CRISPR-Cas9 editing of Elob in mouse models?
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Technical recommendations:
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Design sgRNAs targeting exon 3 (critical for Elongin-B/VHL complex stability) using tools like CHOPCHOP.
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Validate edits via Sanger sequencing and functional assays (e.g., RNA Pol II elongation rates).
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Monitor off-target effects using whole-exome sequencing, referencing reproducibility criteria from electrophysiology protocols .
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Data Integration & Interpretation
How can multi-omics data clarify ELOB’s role in transcription elongation?
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Workflow:
Approach Application Validation ChIP-seq Identify ELOB-bound loci Compare with POLR2A occupancy ATAC-seq Assess chromatin accessibility Correlate with elongation rates Proteomics Map interaction partners (e.g., Elongin-C, VHL) Co-IP with knockout controls
Experimental Reproducibility
What quality controls ensure consistency in ELOB-focused electrophysiology studies?
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Critical steps:
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Adopt Neuropixels probe alignment standards , including stereotaxic coordinates (±50 µm tolerance) for hippocampal recordings.
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Implement session exclusion criteria:
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Minimum 400 behavioral trials
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Probe placement verified via post-hoc histology
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Analyze within-lab vs. cross-lab variability using mixed-effects models .
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Methodological Pitfalls
Why do inconsistencies arise in ELOB knockout phenotype reporting?
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Root-cause analysis:
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Genetic background: Compare C57BL/6J vs. BALB/c strains for viability differences.
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Environmental factors: Standardize housing conditions (e.g., light cycles, diet) per NIH guidelines.
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Endpoint variability: Use longitudinal monitoring instead of single-timepoint assessments.
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Product Science Overview
Elongin B forms a heterodimer with Elongin C (ELOC), and together they serve as the regulatory subunits for the Elongin complex . This complex is a general transcription elongation factor that enhances the transcription process by alleviating transcriptional pausing . The Elongin B/C heterodimer also binds to the “BC-box motif” found in many proteins within the VHL-box and SOCS-box protein families .
Elongin B is also a component of Cullin-RING E3 ubiquitin ligase complexes (CRLs), specifically those based on Cullin-2 (Cul2) and Cullin-5 (Cul5) . These complexes play a central role in targeting cellular proteins for ubiquitination-dependent protein turnover through the 26S proteasome . In these complexes, Elongin B, along with Elongin C, acts as an adapter protein that helps in the assembly of the CRL complex .
The Elongin B/C complex is involved in various biological processes, including the regulation of hypoxia-inducible factors (HIFs). The most well-known CRL2 substrate recognition receptor is the tumor suppressor protein VHL (von Hippel–Lindau), which is mutated in von Hippel–Lindau syndrome, a rare hereditary cancer syndrome . The CRL2 VHL complex-dependent degradation of the α subunits of HIF (HIFα) is a critical function in tumorigenesis .
Recombinant Elongin B (Mouse) is produced using various expression systems, including baculovirus-infected insect cells. It is often used in research to study its role in transcription elongation and ubiquitination processes. The recombinant protein is typically supplied as a solution in HEPES, NaCl, DTT, and Glycerol, and it is stored at -70°C to maintain its stability .
