Related Experiment Video
Updated: Jan 20, 2026
09:31
Amplification, Next-generation Sequencing, and Genomic DNA Mapping of Retroviral Integration Sites
Published on: March 22, 2016
18.3K
iDamage: a method to integrate modified DNA into the yeast genome
Katarzyna H Masłowska1, Luisa Laureti1, Vincent Pagès1
1CRCM: Team DNA Damage and Genome Instability | Aix Marseille Univ, CNRS, INSERM, Institut Paoli-Calmettes, Marseille, France.
Nucleic Acids Research
|August 17, 2019
Summary
Researchers developed a precise method to study DNA repair in yeast. Inhibiting homologous recombination significantly increased error-prone Trans Lesion Synthesis (TLS) pathways for DNA damage tolerance.
Area of Science:
- Molecular Biology
- Genetics
- Biochemistry
Background:
- Living cells possess mechanisms to repair DNA alterations.
- Understanding DNA damage tolerance is crucial for cell survival and preventing mutations.
Purpose of the Study:
- To develop a method for site-specific DNA modification in the yeast genome.
- To investigate the regulation of error-free versus error-prone DNA lesion bypass.
Main Methods:
- Site-specific integration of modified DNA into the Saccharomyces cerevisiae genome using the Cre/lox recombination system.
- Introduction of a single UV lesion into the yeast genome.
- Inhibition of homologous recombination via Rad51 inactivation or ubc13 mutation.
Main Results:
- The developed method allows precise and robust integration of modified DNA at chosen genomic loci.
- Inhibition of homologous recombination strongly favors the use of Trans Lesion Synthesis (TLS) for DNA damage bypass.
- This regulation of DNA damage tolerance was not observable with previous methods.
Conclusions:
- The new method provides a powerful tool for studying DNA replication and repair.
- Cellular regulation balances error-free and error-prone DNA repair pathways.
- Understanding these mechanisms is key to comprehending cellular responses to DNA damage.
Related Concept Videos
02:02Genomics
39.8K
Genomics is the science of genomes: it is the study of all the genetic material of an organism. In humans, the genome consists of information carried in 23 pairs of chromosomes in the nucleus, as well as mitochondrial DNA. In genomics, both coding and non-coding DNA is sequenced and analyzed. Genomics allows a better understanding of all living things, their evolution, and their diversity. It has a myriad of uses: for example, to build phylogenetic trees, to improve productivity and...
39.8K
00:46Genomic DNA in Prokaryotes
48.3K
The genome of most prokaryotic organisms consists of double-stranded DNA organized into one circular chromosome in a region of cytoplasm called the nucleoid. The chromosome is tightly wound, or supercoiled, for efficient storage. Prokaryotes also contain other circular pieces of DNA called plasmids. These plasmids are smaller than the chromosome and often carry genes that confer adaptive functions, such as antibiotic resistance.
Genomic Diversity in Bacteria
Although bacterial genomes are much...
Genomic Diversity in Bacteria
Although bacterial genomes are much...
48.3K
00:58Genomic DNA in Eukaryotes
52.4K
Eukaryotes have large genomes compared to prokaryotes. To fit their genomes into a cell, eukaryotic DNA is packaged extraordinarily tightly inside the nucleus. To achieve this, DNA is tightly wound around proteins called histones, which are packaged into nucleosomes that are joined by linker DNA and coil into chromatin fibers. Additional fibrous proteins further compact the chromatin, which is recognizable as chromosomes during certain phases of cell division.
52.4K
02:36DNA Damage Can Stall the Cell Cycle
3.1K
In response to DNA damage, cells can pause the cell cycle to assess and repair the breaks. However, the cell must check the DNA at certain critical stages during the cell cycle. If the cell cycle pauses before DNA replication, the cells will contain twice the amount of DNA. On the other hand, if cells arrest after DNA replication but before mitosis, they will contain four times the normal amount of DNA. With a host of specialized proteins at their disposal,cells must use the right protein at...
3.1K
02:36DNA Damage can Stall the Cell Cycle
10.0K
In response to DNA damage, cells can pause the cell cycle to assess and repair the breaks. However, the cell must check the DNA at certain critical stages during the cell cycle. If the cell cycle pauses before DNA replication, the cells will contain twice the amount of DNA. On the other hand, if cells arrest after DNA replication but before mitosis, they will contain four times the normal amount of DNA. With a host of specialized proteins at their disposal,cells must use the right protein at...
10.0K
01:28Yeast Signaling
17.2K
Yeasts are single-celled organisms, but unlike bacteria, they are eukaryotes (cells with a nucleus). Cell signaling in yeast is similar to signaling in other eukaryotic cells. A ligand, such as a protein or a small molecule released from a yeast cell, attaches to a receptor on the cell surface. The binding stimulates second-messenger kinases to activate or inactivate transcription factors that further regulate gene expression. Many of the yeast intracellular signaling cascades have similar...
17.2K