From the Guidelines
Diagnosing cytoplasmic DNA presence is typically done through methods such as next-generation sequencing, PCR, and DNAse digestion.
Key Methods
- Next-generation sequencing: to analyze EV-DNA content 1
- PCR: to validate the EV-DNA content 1
- DNAse digestion: to demonstrate the presence of intravesicular DNA, a DNAse digestion step of intact EVs is needed 1 These methods are essential in detecting the presence of DNA in extracellular vesicles (EVs), which can differ from DNA found in apoptotic bodies, especially under stress conditions.
Importance of DNAse Digestion
The use of DNAse digestion is crucial as it helps to distinguish between DNA that is truly intravesicular and DNA that may be adhering to the outside of the EVs. By employing these methods, researchers and clinicians can accurately diagnose the presence of cytoplasmic DNA, which is vital for understanding various biological processes and potential disease mechanisms.
From the Research
Detection Methods
- Fluorescence in situ hybridization (FISH) can be used for the intracellular detection of DNA or RNA molecules, however, RNA FISH can detect cytoplasmic DNA without the need for denaturation 2.
- Whole-genomic amplification (WGA) can be used to detect DNA in blastocoelic fluid, which can be correlated with embryo ploidy and viable pregnancy 3.
Cytoplasmic DNA Dynamics
- Autophagy regulates cytoplasmic dsDNA accumulation and subsequent redistribution into extracellular vesicles 4.
- Accumulated cytoplasmic dsDNAs can activate the STING pathway, reducing leukemia cell viability through reactive oxygen species (ROS) generation 4.
- STING recognition of cytoplasmic DNA instigates cellular defense, including the activation of innate immune and proinflammatory genes 5.
Implications
- The presence of DNA in blastocoelic fluid can be correlated with blastocyst ploidy condition and implantation potential 3.
- Manipulation of cytoplasmic dsDNA dynamics can be a novel and potent therapeutic strategy for myeloid leukemias 4.
- Cytoplasmic membrane-associated transcription of cytoplasmic DNA can occur through a special transcription system involving DNA-dependent RNA polymerase II 6.