About event
Dear Colleagues,
Mendel Center Seminar (Oct 1st) has been canceled due to the illness of the lecturer.
Thank you for your understanding.
The seminars are devoted to topics that develop directions of the research groups of the Mendel Center for Plant Genomics and Proteomics. The speakers are primarily prominent foreign and domestic experts. In addition, some seminars are dedicated to the presentation of research results by internal speakers, mainly postdoctoral fellows and PhD candidates. Seminars are thus a platform for sharing knowledge, establishing cooperation and developing communication skills.
More information
Understanding the three-dimensional organization of chromatin within the nucleus is essential for comprehending gene expression regulation during normal development and disease. Traditional methods of visualizing chromatin structure, such as fluorescence in situ hybridization (FISH), often require complete DNA denaturation, which can disrupt the natural chromatin conformation. CRISPR-FISH, a novel technique, overcomes this limitation by preserving chromatin integrity. It utilizes target-specific crRNA, fluorescence-labeled tracrRNA, and recombinant dCas9 protein to selectively label repetitive DNA sequences in fixed plant and animal nuclei and chromosomes. In this lecture, I introduce CRISPR-CID (CRISPR/dCas9-mediated chromogenic in situ detection), a novel imaging technique that integrates CRISPR technology with chromogenic signal detection. By utilizing horseradish peroxidase, CRISPR-CID enables non-fluorescent labeling of repetitive DNA sequences, making it possible to visualize these sequences with conventional bright-field microscopy. This technique is particularly valuable for educational institutions and outreach programs with limited access to fluorescence microscopes. Additionally, I report improvements to the CRISPR-FISH method aimed at increasing signal intensity by engineering a recombinant dCas9 protein with an ALFA-tag. Finally, we are developing CRISPR-dCas9-based strategies to identify DNA-binding proteins.
