Related Experiment Video
Updated: May 23, 2026
12:16
Robust 3D DNA FISH Using Directly Labeled Probes
Published on: August 15, 2013
A comparative study of cell-free apoptotic and genomic DNA using FISH and massive parallel sequencing
Evgeniy S Morozkin1, Ekaterina M Loseva, Igor V Morozov
1Institute of Chemical Biology and Fundamental Medicine, Siberian Branch, Russian Academy of Sciences, pr. Lavrentieva 8, Novosibirsk, Russian Federation. morozkin@niboch.nsc.ru
Expert Opinion on Biological Therapy
|April 18, 2012
Summary
Apoptosis generates cell-free DNA (cfDNA) enriched with Alu repeats and coding sequences, offering insights into diagnostic targets. This study compared apoptotic and genomic DNA to understand cfDNA generation mechanisms.
Area of Science:
- Molecular Biology
- Genetics
- Biochemistry
Background:
- Circulating DNA (cirDNA) generation mechanisms are crucial for identifying diagnostic molecular targets.
- Apoptosis is a significant source of cirDNA, necessitating detailed study of its characteristics.
Purpose of the Study:
- To conduct an in vitro comparative study of cell-free apoptotic DNA (cm-apoDNA) and genomic DNA (gDNA).
- To elucidate the influence of apoptosis on cirDNA content and composition for diagnostic applications.
Main Methods:
- Isolation of DNA from culture medium of apoptotic human umbilical vein endothelial cells (cm-apoDNA) and gDNA from living cells.
- Analysis of DNA using Fluorescence In Situ Hybridization (FISH) and next-generation sequencing (SOLiD 3 platform).
Main Results:
- FISH revealed overrepresentation of C-positive chromosome regions in cm-apoDNA.
- SOLiD 3 sequencing showed significant enrichment of cm-apoDNA for specific Alu repeats (AluJ, AluS, AluY) and underrepresentation of certain L1 elements (L1MA, L1ME).
- Data indicated non-uniform genomic DNA degradation during apoptosis.
Conclusions:
- Apoptosis leads to non-uniform degradation of genomic DNA, resulting in cell-free apoptotic DNA enriched with Alu repeats and coding sequences.
- These findings highlight the importance of understanding apoptosis-driven cirDNA generation for developing effective diagnostic tools.
Related Concept Videos
02:54Evolutionary Relationships through Genome Comparisons
Genome comparison is one of the excellent ways to interpret the evolutionary relationships between organisms. The basic principle of genome comparison is that if two species share a common feature, it is likely encoded by the DNA sequence conserved between both species. The advent of genome sequencing technologies in the late 20th century enabled scientists to understand the concept of conservation of domains between species and helped them to deduce evolutionary relationships across diverse...
03:00Next-generation Sequencing
The first human genome sequencing project cost $2.7 billion and was declared complete in 2003, after 15 years of international cooperation and collaboration between several research teams and funding agencies. Today, with the advent of next-generation sequencing technologies, the cost and time of sequencing a human genome have dropped over 100 fold.
Next-Generation Sequencing Methods
Although all next-generation methods use different technologies, they all share a set of standard features.
Next-Generation Sequencing Methods
Although all next-generation methods use different technologies, they all share a set of standard features.
01:57Sanger Sequencing
DNA sequencing is a fundamental technique that is routinely used in the biological sciences. This method can be applied to a range of questions at different scales - from the sequencing of a cloned DNA fragment or the study of a mutation in a gene up to whole-genome sequencing. However, despite the widespread use of sequencing today, it was not until 1977 that Fredrick Sanger and his collaborators developed the chain-termination method to decode DNA sequences. It relies on the separation of a...