GO:0042138 meiotic DNA double-strand break formation: Initiation of Meiotic Recombination, Genes, Functions and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:0042138 describes the programmed generation of DNA double-strand breaks (DSBs) at defined hotspots during meiosis I, which initiates meiotic recombination.
• The catalytic core is the SPO11 topoisomerase-like enzyme, which is reconstituted in vitro together with accessory factors to cleave DNA.
• DSB formation is tightly regulated by ATM/Tel1-dependent feedback and by meiotic prophase length, which controls DSB interference and distribution.
• Accessory proteins such as ZmMTOPVIB in maize and splicing regulators BCAS2/hnRNPH1 are required for efficient DSB formation and downstream repair.
• Defects in DSB formation cause meiotic arrest, aneuploidy and infertility, and are linked to cancer predisposition through genome instability.
• CRISPR knockout, point-mutation, knock-in and overexpression models enable causal dissection of DSB formation genes in human cells and model organisms.
Description
Meiotic DNA double-strand break formation (GO:0042138) is the cell cycle process in which double-strand breaks (DSBs) are generated at defined hotspots throughout the genome during meiosis I, initiating meiotic recombination. This process is essential for faithful chromosome segregation and genetic diversity, and its dysregulation is associated with infertility and genome instability. Understanding the molecular players and regulatory logic of DSB formation is therefore central to reproductive biology and cancer research. Recent in vitro reconstitution studies have defined the minimal biochemical machinery required for SPO11-dependent DSB formation, providing a mechanistic framework for this GO term. In parallel, genetic studies in plants and vertebrates have revealed conserved and species-specific accessory factors that control DSB number, timing and distribution. This article integrates authoritative QuickGO annotation with verified PubMed literature to provide a research-grade overview of GO:0042138, its genes, regulation, disease relevance and experimental methods.
meiotic DNA double-strand break formation At A Glance
| GO ID | GO:0042138 |
|---|---|
| GO term | meiotic DNA double-strand break formation |
| Ontology | biological_process |
| Synonym | none |
| Major function | Generation of programmed DNA double-strand breaks at meiotic hotspots to initiate recombination |
| Catalytic core | SPO11 and accessory proteins (e.g., MTOPVIB, TOP6BL) |
| Cell cycle context | Meiosis I prophase |
| Regulatory input | ATM/Tel1-dependent feedback and meiotic prophase length |
| Downstream outcome | Meiotic recombination, synapsis and chromosome segregation |
What Is GO:0042138?
GO:0042138 (meiotic DNA double-strand break formation) is defined as the cell cycle process in which double-strand breaks are generated at defined hotspots throughout the genome during meiosis I, resulting in the initiation of meiotic recombination. In other words, it is the programmed, enzyme-catalyzed creation of DNA breaks that serves as the first committed step of meiotic recombination, rather than a damage response.
Why Is meiotic DNA double-strand break formation Important in Cell Biology?
Meiotic DNA double-strand break formation is important because it is the initiating event of meiotic recombination, which is required for accurate chromosome segregation and genetic diversity. Defects in this process lead to meiotic arrest, aneuploidy and infertility, and impaired DSB formation or repair is associated with genome instability and cancer predisposition. Because DSB formation is tightly regulated by ATM/Tel1 signaling and prophase timing, it also serves as a paradigm for understanding how cells balance break formation with repair capacity.
• Initiates meiotic recombination and is essential for faithful chromosome segregation.
• Generates genetic diversity through crossing over and gene conversion.
• Its failure causes meiotic arrest and infertility.
• Dysregulation is linked to aneuploidy and genome instability.
• ATM/Tel1 feedback controls DSB number and distribution.
• Accessory factors such as ZmMTOPVIB are required for DSB formation in maize.
• Splicing regulators BCAS2 and hnRNPH1 couple DSB repair to synapsis.
• In vitro reconstitution enables biochemical dissection of the core machinery.
• Provides targets for reproductive and cancer research.
• Supports development of CRISPR models for causal gene testing.
What Happens During meiotic DNA double-strand break formation?
Initiation at meiotic hotspots
In simple terms: The cell deliberately cuts its own DNA at specific spots to start recombination.
During meiosis I, DSBs are generated at defined hotspots throughout the genome, initiating meiotic recombination. This programmed breakage is not random damage but a regulated process that marks the start of recombination. Hotspot distribution and DSB number are modulated by meiotic prophase length and Tel1-dependent interference.
SPO11-dependent catalysis
In simple terms: A specialized enzyme called SPO11 makes the cuts in DNA.
The catalytic core of DSB formation is SPO11, a topoisomerase-like enzyme that generates DSBs in vitro when reconstituted with accessory factors. Reconstitution of SPO11-dependent DSB formation has defined the minimal biochemical requirements for this reaction. Accessory proteins such as ZmMTOPVIB are required for DSB formation and bipolar spindle assembly during maize meiosis.
Accessory factors and complex assembly
In simple terms: Helper proteins assemble with SPO11 to make the cutting machinery work.
DSB formation requires accessory factors in addition to SPO11, as shown by in vitro reconstitution and genetic studies. In maize, ZmMTOPVIB enables DSB formation and is also required for bipolar spindle assembly. Splicing regulators BCAS2 and hnRNPH1 orchestrate alternative splicing for DSB repair and synapsis in meiotic prophase I, linking DSB formation to downstream events.
ATM/Tel1-dependent feedback and interference
In simple terms: The cell monitors breaks and adjusts how many more it makes.
ATM/Tel1 provides local and global control of DSB formation, ensuring breaks are distributed appropriately. Meiotic prophase length modulates Tel1-dependent DSB interference, affecting the spacing and number of breaks. This feedback prevents excessive breakage and coordinates DSB formation with repair capacity.
Repair and downstream recombination
In simple terms: After the cut, the cell repairs the break using a template to create genetic exchange.
DSBs generated by GO:0042138 are repaired by homologous recombination, producing DNA synthesis tracts that can be mapped in Arabidopsis thaliana. Repair is coupled to synapsis and chromosome pairing, and defects in this coupling cause meiotic arrest. In heteromorphic sex chromosomes, DSB formation and pairing show specialized patterns, as seen in threespine stickleback.
Key Genes Involved in GO:0042138 meiotic DNA double-strand break formation
The following genes and proteins are experimentally implicated in meiotic DNA double-strand break formation and its regulation.
| Gene | Major Role | Research Relevance |
|---|---|---|
| SPO11 | Catalytic core of DSB formation | Reconstituted in vitro for mechanistic studies |
| MTOPVIB | Accessory factor required for DSB formation | Maize mutant shows defective DSB and spindle assembly |
| ATM | Local and global control of DSB formation | Regulates DSB number and distribution |
| Tel1 | Yeast ATM ortholog controlling DSB interference | Modulates DSB spacing with prophase length |
| BCAS2 | Splicing regulator for DSB repair and synapsis | Required for meiotic prophase I progression |
| hnRNPH1 | Splicing regulator for DSB repair and synapsis | Couples splicing to meiotic DSB repair |
| TOP6BL | Accessory factor for SPO11-dependent DSB formation | Part of reconstituted DSB machinery |
| RAD51 | Recombinase acting downstream of DSBs | Marker of DSB repair tracts |
| DMC1 | Meiotic recombinase | Required for interhomolog repair |
| MRE11 | DSB end processing | Required for repair initiation |
| RAD50 | DSB end processing complex | Part of MRN complex |
| NBS1 | DSB end processing complex | Part of MRN complex |
| H2AX | Histone variant marking DSBs | Used as DSB marker in imaging |
| SYP1 | Synaptonemal complex component | Links DSB repair to synapsis |
| ZIP1 | Synaptonemal complex component | Links DSB repair to synapsis |
| SPO11-1 | Plant SPO11 paralog | DSB formation in Arabidopsis |
| SPO11-2 | Plant SPO11 paralog | DSB formation in Arabidopsis |
How Is meiotic DNA double-strand break formation Regulated?
DSB formation is regulated by ATM/Tel1-dependent feedback, which provides local and global control of break number and distribution. Meiotic prophase length modulates Tel1-dependent DSB interference, so timing of prophase progression directly affects DSB spacing. Splicing regulators BCAS2 and hnRNPH1 influence DSB repair and synapsis, indicating that post-transcriptional regulation is part of the control network. Accessory factors such as ZmMTOPVIB are required for DSB formation and are also linked to spindle assembly, suggesting coordination between DSB formation and meiotic progression.
meiotic DNA double-strand break formation and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| ATM | Genome instability and cancer predisposition | ATM knockout cell line |
| BCAS2 | Meiotic arrest and infertility | BCAS2 knockout mouse or cell model |
| hnRNPH1 | Meiotic prophase I defects | hnRNPH1 knockdown/knockout |
| SPO11 | Infertility and meiotic failure | SPO11 knockout mouse |
| MTOPVIB | Meiotic DSB and spindle defects | Maize mutant |
Infertility and meiotic arrest
Defects in meiotic DNA double-strand break formation cause meiotic arrest and are associated with infertility, as DSBs are required to initiate recombination and synapsis. Splicing regulators such as BCAS2 and hnRNPH1 are required for DSB repair and synapsis in meiotic prophase I, and their loss impairs meiotic progression.
Cancer and genome instability
ATM controls DSB formation locally and globally, and its dysfunction is linked to genome instability and cancer predisposition. Impaired DSB formation or repair can lead to aneuploidy and chromosomal rearrangements, which are hallmarks of cancer.
Aneuploidy and reproductive disorders
Failure to form or repair meiotic DSBs leads to aneuploidy, a major cause of miscarriage and reproductive disorders. Studies in heteromorphic sex chromosomes show that DSB formation and pairing are specialized, with implications for sex chromosome disorders.
From meiotic DNA double-strand break formation-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Is a candidate gene required for DSB formation? | CRISPR knockout cell line or organism |
| Does a specific residue control SPO11 catalysis? | Point-mutation knock-in |
| Where and when is the protein expressed? | Tagged knock-in with fluorescent reporter |
| Does overexpression alter DSB number? | Overexpression cell model |
| Which genes modify DSB distribution? | CRISPR library screening |
| How do splicing factors affect DSB repair? | Knockout of BCAS2/hnRNPH1 |
How to Study the meiotic DNA double-strand break formation Process
| Method | What It Measures | Typical Application |
|---|---|---|
| DSB mapping by sequencing | Genome-wide DSB hotspots | Identify break sites |
| In vitro reconstitution | Biochemical DSB formation | Test SPO11 mutants |
| H2AX foci imaging | DSB formation and repair | Cytological assessment |
| Recombination frequency | Genetic crossing over | Mutant analysis |
| RNA-seq | Alternative splicing of DSB genes | BCAS2/hnRNPH1 studies |
| CRISPR knockout | Gene requirement for DSB formation | Functional screens |
| CRISPR library screening | Modifiers of DSB distribution | Genome-wide discovery |
| Prophase timing analysis | Tel1-dependent interference | DSB spacing studies |
DSB mapping and sequencing
DSB formation can be mapped genome-wide by sequencing repair intermediates, as demonstrated by meiotic DSB repair DNA synthesis tracts in Arabidopsis thaliana. This approach identifies hotspots and quantifies break frequency.
In vitro reconstitution
In vitro reconstitution of meiotic DSB formation using purified SPO11 and accessory factors defines the minimal machinery and allows biochemical dissection of catalysis. This method is powerful for testing point mutations and cofactor requirements.
Genetic and cytological assays
DSB formation is assessed genetically by recombination frequency and cytologically by markers such as H2AX foci. Mutants in ZmMTOPVIB show defective DSB formation and spindle assembly in maize.
Splicing and transcript analysis
Alternative splicing of DSB repair genes can be analyzed by RNA-seq, as shown for BCAS2 and hnRNPH1 in meiotic prophase I. This links post-transcriptional regulation to DSB formation and repair.
How CRISPR Can Be Used to Study GO:0042138 meiotic DNA double-strand break formation
Knockout
CRISPR knockout of candidate genes such as SPO11, MTOPVIB, BCAS2 or hnRNPH1 can test their requirement for meiotic DSB formation and downstream repair. Knockout models are also used to assess meiotic arrest and aneuploidy phenotypes.
Point Mutation
Point-mutation knock-in of catalytic residues in SPO11 or accessory factors allows separation of DSB formation from other functions. Such models are valuable for testing biochemical predictions from in vitro reconstitution.
Knock-in
Tagged knock-in of DSB proteins enables live imaging and mapping of DSB sites in cells and organisms. Knock-in of reporter cassettes can also quantify DSB formation at defined hotspots.
Overexpression
Overexpression of DSB factors or regulators such as ATM can test whether increased dosage alters DSB number or distribution. Overexpression models complement loss-of-function studies to define sufficiency.
How EDITGENE Supports meiotic DNA double-strand break formation Research
Researchers studying meiotic DNA double-strand break formation-related genes often need to determine whether a candidate gene is causally involved in DSB formation, repair or meiotic progression. EDITGENE provides CRISPR-based cell models and screening services to test these hypotheses with publication-ready rigor.
Contact EDITGENE today to design your custom CRISPR model for meiotic DNA double-strand break formation research.
Related Products
| Product name | Cat.No. | Species | Gene ID | |
|---|---|---|---|---|
| MRE11 Knockout HEK293 Cell Line | EDJ-KQ3753 | Human | 4361 | Details Get a Quote |
| M1AP Knockout HEK293 Cell Line | EDJ-KQ9260 | Human | 130951 | Details Get a Quote |
| HORMAD1 Knockout HEK293 Cell Line | EDJ-KQ9967 | Human | 84072 | Details Get a Quote |
| TOP6BL Knockout HEK293 Cell Line | EDJ-KQ12653 | Human | 79703 | Details Get a Quote |
| IHO1 Knockout HEK293 Cell Line | EDJ-KQ13820 | Human | 339834 | Details Get a Quote |
| MEI4 Knockout HEK293 Cell Line | EDJ-KQ14230 | Human | 101928601 | Details Get a Quote |
| SHOC1 Knockout HEK293 Cell Line | EDJ-KQ14470 | Human | 158401 | Details Get a Quote |
| REC114 Knockout HEK293 Cell Line | EDJ-KQ15027 | Human | 283677 | Details Get a Quote |
| SPO11 Knockout HEK293 Cell Line | EDJ-KQ15489 | Human | 23626 | Details Get a Quote |
| TEX11 Knockout HEK293 Cell Line | EDJ-KQ15690 | Human | 56159 | Details Get a Quote |
| SHOC1 Knockout HCT 116 Cell Line | EDJ-KQ45938 | Human | 158401 | Details Get a Quote |
| MRE11 Knockout A-549 Cell Line | EDJ-KQ25827 | Human | 4361 | Details Get a Quote |
| MRE11 Knockout HCT 116 Cell Line | EDJ-KQ25828 | Human | 4361 | Details Get a Quote |
| MRE11 Knockout HeLa Cell Line | EDJ-KQ25829 | Human | 4361 | Details Get a Quote |
| M1AP Knockout HCT 116 Cell Line | EDJ-KQ35863 | Human | 130951 | Details Get a Quote |
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Frequently Asked Questions About meiotic DNA double-strand break formation
What is meiotic DNA double-strand break formation?
It is the process (GO:0042138) in which double-strand breaks are generated at defined hotspots during meiosis I to initiate meiotic recombination.
What genes are involved in meiotic DNA double-strand break formation?
Key genes include SPO11, MTOPVIB, ATM, Tel1, BCAS2, hnRNPH1 and accessory factors such as TOP6BL.
Why is SPO11 important for meiotic DSB formation?
SPO11 is the catalytic core that generates DSBs, as shown by in vitro reconstitution of SPO11-dependent DSB formation.
How is meiotic DSB formation regulated?
It is regulated by ATM/Tel1-dependent feedback and by meiotic prophase length, which controls DSB interference.
What happens if meiotic DSB formation fails?
Failure causes meiotic arrest, aneuploidy and infertility, and is linked to genome instability.
How can I study meiotic DSB formation in the lab?
Methods include DSB mapping by sequencing, in vitro reconstitution, H2AX foci imaging and CRISPR knockout models.
What is the role of ZmMTOPVIB in maize meiosis?
ZmMTOPVIB enables DSB formation and bipolar spindle assembly during maize meiosis.
How do BCAS2 and hnRNPH1 affect meiotic DSBs?
They orchestrate alternative splicing for DSB repair and synapsis in meiotic prophase I.
Can CRISPR be used to study meiotic DSB genes?
Yes, CRISPR knockout, point-mutation, knock-in and overexpression models are used to test DSB gene function.
What diseases are linked to meiotic DSB formation defects?
Infertility, aneuploidy and cancer predisposition are linked to defects in DSB formation and repair.
Conclusion
GO:0042138 (meiotic DNA double-strand break formation) is the programmed initiation of meiotic recombination, driven by SPO11 and accessory factors and regulated by ATM/Tel1 feedback and prophase timing. Its correct execution is essential for fertility and genome stability, and its dysregulation contributes to aneuploidy and cancer. CRISPR-based knockout, point-mutation, knock-in and overexpression models, combined with DSB mapping and in vitro reconstitution, provide powerful tools to dissect this process.
References
- 1. Hernández Sánchez-Rebato M et al.. 2024. Meiotic double-strand break repair DNA synthesis tracts in Arabidopsis thaliana.. PLoS Genet 20(7):e1011197 PMID: 39012914
- 2. Tang X et al.. 2025. In vitro reconstitution of meiotic DNA double-strand-break formation.. Nature 639(8055):800-807 PMID: 39972125
- 3. López Ruiz LM et al.. 2024. Meiotic prophase length modulates Tel1-dependent DNA double-strand break interference.. PLoS Genet 20(3):e1011140 PMID: 38427688
- 4. Lukaszewicz A et al.. 2018. Control of meiotic double-strand-break formation by ATM: local and global views.. Cell Cycle 17(10):1155-1172 PMID: 29963942
- 5. Jing JL et al.. 2020. ZmMTOPVIB Enables DNA Double-Strand Break Formation and Bipolar Spindle Assembly during Maize Meiosis.. Plant Physiol 184(4):1811-1822 PMID: 33077613
- 6. Sun L et al.. 2024. BCAS2 and hnRNPH1 orchestrate alternative splicing for DNA double-strand break repair and synapsis in meiotic prophase I.. Cell Mol Life Sci 81(1):449 PMID: 39520542
- 7. Zheng Z et al.. 2025. Reconstitution of SPO11-dependent double-strand break formation.. Nature 639(8055):784-791 PMID: 39972129
- 8. Nath S et al.. 2022. Meiotic pairing and double-strand break formation along the heteromorphic threespine stickleback sex chromosomes.. Chromosome Res 30(4):429-442 PMID: 35635635
