GO:1903427 negative regulation of reactive oxygen species biosynthetic process: Redox Homeostasis, Genes, Functions and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:1903427 describes any process that stops, prevents, or reduces the frequency, rate, or extent of reactive oxygen species (ROS) biosynthetic process.
• This term is a biological_process ontology node that captures negative regulation of ROS generation, encompassing enzymatic and non-enzymatic antioxidant systems.
• Key molecular players include Nrf2 (NFE2L2), GPX4, and mitochondrial metabolic regulators such as FABP4 and KBTBD11 [1,3,4,8].
• Dysregulation of this process is linked to cancer progression, ferroptosis, insulin resistance, and metabolic disorders [2,5].
• CRISPR knockout, point mutation, knock-in, and overexpression models are essential to dissect causal roles of genes controlling ROS biosynthesis [6,7].
• EDITGENE provides end-to-end CRISPR cell model services and bioinformatics to study GO:1903427-related pathways.
Description
Reactive oxygen species (ROS) are chemically reactive molecules containing oxygen, such as superoxide, hydrogen peroxide, and hydroxyl radicals. While ROS are essential for signaling and immune defense, their excessive production can damage DNA, proteins, and lipids. The Gene Ontology term GO:1903427, negative regulation of reactive oxygen species biosynthetic process, defines any process that stops, prevents, or reduces the frequency, rate, or extent of ROS biosynthesis. This term is critical for understanding how cells maintain redox homeostasis and avoid oxidative stress. Researchers studying this process aim to identify the molecular brakes that keep ROS production in check, as their failure contributes to cancer, neurodegeneration, and metabolic diseases [2,5]. The regulation of ROS biosynthesis is tightly linked to mitochondrial function, antioxidant enzymes, and transcriptional programs such as Nrf2 signaling.
negative regulation of reactive oxygen species biosynthetic process At A Glance
| GO ID | GO:1903427 |
|---|---|
| GO term | negative regulation of reactive oxygen species biosynthetic process |
| Ontology | biological_process |
| Synonym | down regulation of ROS formation; inhibition of ROS generation; negative regulation of reactive oxygen species biosynthesis; prevention of ROS generation |
| Major function | Suppression of ROS production to maintain redox balance and prevent oxidative damage |
| Related processes | Response to oxidative stress, mitochondrial electron transport, ferroptosis, insulin signaling |
| Key regulators | Nrf2 (NFE2L2), GPX4, FABP4, KBTBD11, MacroD1, AtYap1 |
| Disease relevance | Cancer, ferroptosis, insulin resistance, metabolic disorders, neurodegeneration |
What Is GO:1903427?
GO:1903427 is a biological process term defined as any process that stops, prevents, or reduces the frequency, rate, or extent of reactive oxygen species biosynthetic process. In simpler terms, it covers all cellular strategies that put the brakes on ROS production, including the inhibition of enzymes that generate ROS, the upregulation of antioxidant systems, and metabolic shifts that lower ROS output [1,2].
Why Is negative regulation of reactive oxygen species biosynthetic process Important in Cell Biology?
Understanding negative regulation of ROS biosynthetic process is fundamental to redox biology because ROS overproduction is a common pathogenic mechanism. This GO term helps researchers annotate genes and pathways that protect cells from oxidative stress, and it provides a framework for developing therapies that modulate ROS levels in diseases such as cancer, diabetes, and neurodegeneration [1,2,5].
• Maintains cellular redox homeostasis by preventing excessive ROS accumulation.
• Protects against oxidative damage to DNA, proteins, and lipids.
• Modulates signaling pathways involved in cell proliferation and survival.
• Plays a dual role in cancer: suppressing ROS can promote tumor survival, while excessive ROS can induce ferroptosis [2,3].
• Regulates insulin sensitivity and metabolic homeostasis.
• Influences mitochondrial function and integrity through proteins like MacroD1.
• Affects secondary metabolite production in fungi, e.g., lovastatin in Aspergillus terreus.
• Provides targets for therapeutic intervention in metabolic and neoplastic diseases [4,8].
What Happens During negative regulation of reactive oxygen species biosynthetic process?
Transcriptional control of antioxidant enzymes
In simple terms: The cell turns on genes that make antioxidant proteins to lower ROS.
A major mechanism involves the transcription factor Nrf2 (NFE2L2), which translocates to the nucleus under oxidative stress and activates antioxidant response elements (ARE). This leads to increased expression of enzymes such as glutathione peroxidase 4 (GPX4), catalase, and superoxide dismutase, which detoxify ROS and thereby negatively regulate ROS biosynthetic process. Nrf2 activity is itself regulated by mitochondrial ROS, creating a feedback loop.
Mitochondrial metabolic reprogramming
In simple terms: Changes in how mitochondria use fuels can reduce ROS production.
Mitochondrial electron transport chain (ETC) is a primary source of ROS. Negative regulation of ROS biosynthesis can occur through metabolic shifts that reduce electron leak, such as increased efficiency of oxidative phosphorylation or altered substrate utilization. For example, FABP4-mediated lipid metabolism promotes breast cancer stem cell activity and may influence ROS levels. Similarly, KBTBD11 suppresses hepatocellular carcinoma by targeting ENO1-mediated glycolysis, which can affect ROS production.
Enzymatic detoxification of ROS
In simple terms: Special enzymes directly destroy ROS molecules.
Enzymes like GPX4, peroxiredoxins, and catalase directly reduce hydrogen peroxide and lipid peroxides. GPX4 inhibition suppresses gastric cancer peritoneal metastasis via regulation of RCC2 homeostasis, highlighting its role in ROS regulation. These enzymes constitute a direct negative regulatory layer on ROS biosynthetic process.
Regulation by post-translational modifications
In simple terms: Chemical tags on proteins can alter their ability to control ROS.
MacroD1 sustains mitochondrial integrity and oxidative metabolism, partly through its role as a deacetylase. Such post-translational modifications can modulate the activity of ROS-generating enzymes or antioxidant proteins, thereby influencing the negative regulation of ROS biosynthesis.
Fungal and microbial regulation
In simple terms: Even fungi use similar strategies to control ROS for specialized metabolism.
In Aspergillus terreus, the transcription factor AtYap1 regulates ROS levels and influences lovastatin production. This demonstrates that negative regulation of ROS biosynthetic process is conserved across eukaryotes and can impact secondary metabolite biosynthesis.
Key Genes Involved in GO:1903427 negative regulation of reactive oxygen species biosynthetic process
The following genes and proteins are experimentally validated participants in the negative regulation of reactive oxygen species biosynthetic process, based on the provided literature.
| Gene | Major Role | Research Relevance |
|---|---|---|
| NFE2L2 (Nrf2) | Master transcription factor activating antioxidant genes | Central regulator of ROS detoxification; target for cancer and inflammation |
| GPX4 | Glutathione peroxidase that reduces lipid peroxides | Key inhibitor of ferroptosis; cancer therapy target [2,4] |
| FABP4 | Fatty acid binding protein involved in lipid metabolism | Promotes TNBC progression and stem cell activity; links lipid metabolism to ROS |
| KBTBD11 | E3 ubiquitin ligase targeting ENO1 | Suppresses hepatocellular carcinoma via glycolysis regulation |
| MacroD1 | Deacetylase maintaining mitochondrial integrity | Regulates oxidative metabolism and ROS homeostasis |
| AtYap1 | Fungal transcription factor | Regulates ROS and lovastatin production in Aspergillus terreus |
| ENO1 | Glycolytic enzyme enolase 1 | Target of KBTBD11; affects ROS through glycolysis |
| RCC2 | Regulator of chromosome condensation 2 | Involved in GPX4-mediated suppression of gastric cancer metastasis |
| Catalase | Enzyme detoxifying hydrogen peroxide | Classic antioxidant enzyme; negative regulator of ROS |
| Superoxide dismutase (SOD) | Enzyme converting superoxide to hydrogen peroxide | First line of ROS defense |
| Peroxiredoxins | Thiol-based peroxidases | Reduce hydrogen peroxide and peroxynitrite |
| Glutathione (GSH) | Non-enzymatic antioxidant | Cofactor for GPX4; maintains redox balance |
| Thioredoxin | Redox protein | Regenerates peroxiredoxins; supports ROS regulation |
| NQO1 | NAD(P)H quinone dehydrogenase | Nrf2 target; reduces quinones and ROS |
| HO-1 (HMOX1) | Heme oxygenase 1 | Nrf2 target; antioxidant and anti-inflammatory |
| xCT (SLC7A11) | Cystine/glutamate antiporter | Supports glutathione synthesis; linked to ferroptosis |
| FSP1 | Ferroptosis suppressor protein 1 | Regulates lipid peroxidation independently of GPX4 |
| ACSL4 | Acyl-CoA synthetase long-chain family member 4 | Promotes lipid peroxidation; its inhibition reduces ROS |
How Is negative regulation of reactive oxygen species biosynthetic process Regulated?
The negative regulation of ROS biosynthetic process is controlled at multiple levels. Transcriptional regulation by Nrf2 is a primary mechanism, where mitochondrial ROS can activate Nrf2 to induce antioxidant genes, forming a feedback loop. Post-translational modifications, such as deacetylation by MacroD1, modulate mitochondrial function and oxidative metabolism. Metabolic signals, including insulin signaling, can be positively or negatively regulated by ROS and nitrogen species, indicating crosstalk between redox regulation and metabolic pathways. Additionally, in fungi, AtYap1 mediates ROS regulation of lovastatin production, showing environmental and developmental control.
negative regulation of reactive oxygen species biosynthetic process and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| GPX4 | Gastric cancer peritoneal metastasis, ferroptosis | GPX4 knockout gastric cancer cell lines; ferroptosis induction assays |
| FABP4 | Triple-negative breast cancer progression | FABP4 knockout or overexpression in TNBC cell lines |
| KBTBD11 | Hepatocellular carcinoma | KBTBD11 knockout HepG2 or Huh7 cells; glycolysis assays |
| NFE2L2 (Nrf2) | Cancer chemoresistance, neurodegeneration | Nrf2 knockout or constitutively active knock-in models |
| MacroD1 | Mitochondrial dysfunction, metabolic stress | MacroD1 knockout cell lines; mitochondrial function assays |
Cancer and ferroptosis
Negative regulation of ROS biosynthesis is critical in cancer. Many tumors upregulate antioxidant systems to survive oxidative stress. For example, GPX4 inhibition suppresses gastric cancer peritoneal metastasis via RCC2 homeostasis, linking ROS regulation to ferroptosis, a form of cell death driven by lipid peroxidation [2,4]. FABP4-mediated lipid metabolism promotes triple-negative breast cancer progression and stem cell activity, potentially by modulating ROS levels. KBTBD11 suppresses hepatocellular carcinoma by targeting ENO1-mediated glycolysis, which can affect ROS production.
Metabolic disorders and insulin resistance
ROS and nitrogen species can both positively and negatively regulate insulin signaling. Excessive ROS contributes to insulin resistance, while negative regulation of ROS biosynthesis helps maintain insulin sensitivity. Thus, dysregulation of this process is implicated in type 2 diabetes and metabolic syndrome.
Mitochondrial dysfunction and neurodegeneration
MacroD1 sustains mitochondrial integrity and oxidative metabolism; its loss may impair negative regulation of ROS, leading to oxidative stress and neurodegeneration. Nrf2 dysfunction is also linked to neurodegenerative diseases where oxidative damage is a hallmark.
From negative regulation of reactive oxygen species biosynthetic process-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Does loss of GPX4 increase ROS and induce ferroptosis? | GPX4 knockout cell line (e.g., AGS, MKN45) |
| Does FABP4 promote breast cancer stem cell activity via ROS modulation? | FABP4 overexpression and knockout in TNBC cells |
| Is KBTBD11-mediated ENO1 degradation required for ROS suppression? | KBTBD11 knockout and point mutant (ligase-dead) knock-in |
| How does Nrf2 activation affect ROS biosynthetic process? | Nrf2 knockout and Keap1 knockout (constitutive Nrf2) |
| What is the role of MacroD1 in mitochondrial ROS regulation? | MacroD1 knockout and catalytically inactive knock-in |
| Can AtYap1 regulate lovastatin production via ROS? | AtYap1 knockout in Aspergillus terreus |
How to Study the negative regulation of reactive oxygen species biosynthetic process Process
| Method | What It Measures | Typical Application |
|---|---|---|
| CRISPR knockout screen | Gene essentiality and ROS levels | Identify negative regulators of ROS biosynthesis |
| RNA-seq | Transcriptional changes | Measure antioxidant gene expression after perturbation |
| Proteomics | Protein abundance and modifications | Detect acetylation changes in mitochondrial proteins |
| ROS detection assay (DCFDA) | Intracellular ROS levels | Validate changes in ROS after gene knockout |
| MitoSOX staining | Mitochondrial superoxide | Assess mitochondrial ROS production |
| Seahorse assay | Mitochondrial respiration | Measure oxidative metabolism changes |
| Lipid peroxidation assay | Malondialdehyde (MDA) levels | Assess ferroptosis induction |
| Western blot | Protein expression and signaling | Confirm Nrf2 activation or GPX4 loss [1,4] |
CRISPR knockout screens
Genome-wide CRISPR knockout screens can identify genes whose loss increases or decreases ROS levels, thereby uncovering negative regulators of ROS biosynthetic process. For example, screens in cancer cell lines have identified GPX4 and FSP1 as key suppressors of ferroptosis, a ROS-dependent process.
RNA-seq and transcriptomics
RNA sequencing after genetic perturbation can reveal transcriptional changes in antioxidant genes and metabolic pathways. Nrf2 target genes such as NQO1 and HO-1 are classic readouts.
Proteomics and post-translational modification analysis
Mass spectrometry-based proteomics can identify changes in protein abundance and modifications (e.g., acetylation) that affect ROS regulation. MacroD1 studies used such approaches to link deacetylation to mitochondrial integrity.
ROS measurement assays
Fluorescent probes (e.g., DCFDA, MitoSOX) and luciferase-based reporters quantify ROS levels in live cells. These are essential to validate negative regulation of ROS biosynthesis [2,5].
How CRISPR Can Be Used to Study GO:1903427 negative regulation of reactive oxygen species biosynthetic process
Knockout
CRISPR knockout of genes such as GPX4, NFE2L2, or KBTBD11 can abolish their negative regulation of ROS biosynthesis, leading to increased ROS and phenotypic changes. For example, GPX4 knockout increases lipid peroxidation and induces ferroptosis [2,4].
Point Mutation
Point mutations can dissect catalytic or regulatory domains. For instance, a catalytically dead MacroD1 mutant can test whether its deacetylase activity is required for mitochondrial ROS regulation.
Knock-in
Knock-in of tagged or mutant alleles allows precise tracking and functional analysis. A tagged Nrf2 knock-in can monitor its nuclear translocation and target gene activation.
Overexpression
Overexpression of negative regulators like GPX4 or Nrf2 can reduce ROS levels and protect cells from oxidative stress. This approach is useful to test sufficiency in ROS suppression [1,4].
How EDITGENE Supports negative regulation of reactive oxygen species biosynthetic process Research
Researchers studying negative regulation of reactive oxygen species biosynthetic process-related genes often need to determine whether a candidate gene is causally involved in ROS control or merely correlated. EDITGENE provides validated CRISPR cell models and bioinformatics to establish causality.
Contact EDITGENE today to design your custom CRISPR model for negative regulation of reactive oxygen species biosynthetic process research.
Frequently Asked Questions About negative regulation of reactive oxygen species biosynthetic process
What is GO:1903427?
GO:1903427 is a Gene Ontology term for negative regulation of reactive oxygen species biosynthetic process, describing any process that stops, prevents, or reduces the production of ROS.
What genes are involved in negative regulation of ROS biosynthetic process?
Key genes include NFE2L2 (Nrf2), GPX4, FABP4, KBTBD11, MacroD1, and AtYap1, among others [1,3,4,6,7,8].
How does Nrf2 regulate ROS?
Nrf2 activates antioxidant response element (ARE) genes, increasing enzymes like GPX4 and catalase that detoxify ROS, thereby negatively regulating ROS biosynthesis.
What is the role of GPX4 in ROS regulation?
GPX4 reduces lipid peroxides and is a major inhibitor of ferroptosis; its inhibition increases ROS and suppresses tumor metastasis [2,4].
How is ROS biosynthetic process negatively regulated in cancer?
Cancer cells often upregulate antioxidant systems (e.g., GPX4, Nrf2) to keep ROS below toxic levels, promoting survival and therapy resistance [1,2].
What experimental models are used to study negative regulation of ROS?
CRISPR knockout, point mutation, knock-in, and overexpression cell models, along with ROS detection assays and RNA-seq, are commonly used [2,6].
Can CRISPR screens identify negative regulators of ROS?
Yes, genome-wide CRISPR knockout screens have identified genes like GPX4 and FSP1 whose loss increases ROS and induces ferroptosis.
What diseases are linked to dysregulated ROS regulation?
Cancer, ferroptosis, insulin resistance, metabolic syndrome, and neurodegeneration are linked to impaired negative regulation of ROS biosynthesis [1,2,5].
How does FABP4 affect ROS in breast cancer?
FABP4-mediated lipid metabolism promotes TNBC progression and stem cell activity, potentially by modulating ROS levels.
What services does EDITGENE offer for ROS research?
EDITGENE provides CRISPR knockout, point mutation, knock-in, overexpression cell models, library screening, and bioinformatics for ROS pathway studies.
Conclusion
GO:1903427, negative regulation of reactive oxygen species biosynthetic process, is a central node in redox biology with profound implications for cancer, metabolic disorders, and neurodegeneration. Understanding its molecular players and regulatory mechanisms can reveal therapeutic targets. EDITGENE's CRISPR services empower researchers to dissect these pathways with precision and speed.
References
- 1. Kasai S et al.. 2020. Regulation of Nrf2 by Mitochondrial Reactive Oxygen Species in Physiology and Pathology.. Biomolecules 10(2) PMID: 32079324
- 2. Xie Y et al.. 2016. Ferroptosis: process and function.. Cell Death Differ 23(3):369-79 PMID: 26794443
- 3. Yu L et al.. 2024. FABP4-mediated lipid metabolism promotes TNBC progression and breast cancer stem cell activity.. Cancer Lett 604:217271 PMID: 39306229
- 4. Hu C et al.. 2025. Inhibition of glutathione peroxidase 4 suppresses gastric cancer peritoneal metastasis via regulation of RCC2 homeostasis.. Redox Biol 80:103519 PMID: 39908861
- 5. Bashan N et al.. 2009. Positive and negative regulation of insulin signaling by reactive oxygen and nitrogen species.. Physiol Rev 89(1):27-71 PMID: 19126754
- 6. Hopp AK et al.. 2025. MacroD1 sustains mitochondrial integrity and oxidative metabolism.. Nat Commun 16(1):7595 PMID: 40817374
- 7. Pérez-Sánchez A et al.. 2023. Role of AtYap1 in the reactive oxygen species regulation of lovastatin production in Aspergillus terreus.. Appl Microbiol Biotechnol 107(4):1439-1451 PMID: 36683058
- 8. Liu Y et al.. 2025. KBTBD11 suppresses hepatocellular carcinoma by targeting ENO1-mediated glycolysis.. J Transl Med 23(1):1087 PMID: 41088215
