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360° Holistic CRISPR:
CRISPR is now being applied in many model and non-model organisms, including humans, and this technique is attracting more and more attention. During this workshop, where we will talk about many theoretical and practical details about CRISPR technology, which is the most popular invention of recent times, I will include CRISPR bioinformatics applications, which is the most difficult part.
Training Content:
- Overview of the CRISPR Cas9 system
- Comparison of CRISPR with other gene editing techniques
- Designing CRISPR in model and non-model organisms
- Target gene region identification
- Methods for designing and testing gRNA
- Microinjection strategies
- Off-target elimination strategies
- CRISPR analysis by molecular methods
- General review and Question and Answer Day
Training Name: CRISPR 360
Training Objective: Overview of CRISPR techniques, choosing the right gRNA and Cas9, learning microinjection and mutant breeding techniques, designing CRISPR in non-model organisms, mutation detection and analysis.
Who Can Attend: Life science researchers interested in gene editing
Training Topics : Designing a CRISPR experiment from start to finish, designing gRNA, genotype analysis
Trainer: Dr. Özlem Gönülkırmaz Çançalar
Training Date: July 8-12, 2024
Mode of Workshop: Online
Sessions and Training Hours: Weekdays 20:00-22:00
- 1st Day
Session 1 (40 min)
- Overview of the CRISPR Cas9 system
- Comparison of CRISPR with other gene editing techniquesDesigning CRISPR in model and non-model organisms
15 minutes break
Session 2 (40 min)
- Target gene region identification
- Detection of target gene region on splice variants
- SNV (Single Nucleotide Variation) search using IGV (Integrated Genome Viewer)
15 minutes break
Session 3 (40 min)
- Designing primers for the targeted gene on NCBI (Primer Blast usage)
- 2nd Day
Session 1 (40 min)
- Selection of the CRISPR technique to be used
- General guidelines for proper gRNA design and overview of internet-based gRNA design programs
- Designing gRNA for a target gene region
15 minutes break
Session 2 (40 min)
- Geneious Prime program installation and first look (account activation)
- Gene alignment and CRISPR annotation in Geneious Prime
15 minutes break
- Session 3 (40 min)Methods for testing gRNA
- Mutant transcript review (on Geneious Prime)
- 3rd Day
Session 1 (40 min)
- Transfer of engineered gRNA and Cas9 into the organism: Microinjection strategiesOff-target elimination strategies
15 minutes break
Session 2 (40 min)
- Galaxy web server introduction and whole genome off-target scanning
15 minutes break
Session 3 (40 min)
- Whole genome off-target screening in progress
- 4th Day
Session 1 (40 min)
- Molecular methods for CRISPR analysis
- HRMA (High Resolution Melt Analysis) analysis with sampling and mutant identification
15 minutes break
Session 2 (40 min)
- Analysis of sanger sequence data in Geneious Prime
15 minutes break
Session 3 (40 min)
- Genotype evaluation, Indel (Insertion-Deletion) and mosaic distinction
- 5th Day
General review and Q&A day.
End of training evaluation test
Education fee 3.800₺
(VAT 20% included)
Briefly About Dr Ozlem
Bees play a vital role as a crucial part of our ecosystems and are essential for the sustainability of agriculture. Ozlem Gonulkirmaz Cancalar’s focal point during her Ph.D. has been to generate mutant lines in bumble bees to investigate the behavioral influence of canonical circadian genes. Thanks to the prestigious Marie Sklodowska-Curie program, she received advanced training in renowned labs in Würzburg, Germany, and Ceske Budejovice, Czech Republic. In these labs, she honed her skills in immunostaining and developing CRISPR techniques in different insect species, such as Drosophila and Firebug.
With these skills, Ozlem successfully developed a CRISPR/Cas9 system for the non-model organism Bombus terrestris, creating a clock mutant bee for the first time. This groundbreaking project required significant resilience and perseverance, as it was the first instance of gene editing in a bumble bee.
Starting this fall, Ozlem will be joining the University of Würzburg in Germany as a postdoctoral researcher. While she has developed expertise in molecular and behavioral disciplines, her future focus will be primarily on neuronal mechanisms.
The Marie Sklodowska-Curie grant is a highly esteemed honor, providing exceptional opportunities for researchers to advance their careers through international training and collaboration. This grant is known for its competitive nature, recognizing and supporting only the most promising researchers across Europe. Being awarded this grant not only signifies excellence in research but also opens doors to unparalleled academic and professional development, making it a highly prestigious achievement in the scientific community.
First gene editing and circadian rhythm with CRISPR in bumblebees
Bees play an indispensable role in our ecosystems and are vital for the sustainability of agriculture. However, bee populations face numerous threats. Innovative gene-editing techniques like CRISPR offer promising solutions for enhancing bee health and adaptability.
Circadian rhythm regulates many physiological and behavioral processes in various organisms. In bees, it directly influences time management for optimal flower yield, sun-based navigation, social synchronization, and task organization within the colony. The bumble bee, Bombus terrestris, serves as an excellent model for studying the impact of clock genes on behavior due to its small colonies and relative ease of laboratory maintenance compared to honey bees.
In her doctoral research, she was the first to characterize a set of complex behaviors regulated by the circadian rhythm in bumble bees. Additionally, She pioneered the use of the CRISPR/Cas9 gene-editing system in bumble bees to mutate a clock gene.
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