Recycling has always been a smart idea, and nature has its own processes to ensure that waste is kept to a minimum. As Professor Ohsumi discovered, autophagy is the cells way of degrading and recycling cellular components, allowing it to adapt to nutritional deficiency or other environmental influences. Professor Yoshinori Ohsumi, honorary professor and leader of the Cell Biology Unit at the Tokyo Institute of Technology, has been studying autophagy for 27 years. This year's Nobel Laureate discovered and elucidated mechanisms underlying autophagy, according to the Press Release from The Nobel Assembly at Karolinska Institutet.
In more detail, autophagy describes the pathway where by components of the cytoplasm are isolated into double-membrane–bound autophagosomes, and the processes involved in transporting these to the lysosome for degradation. The core genes involved in the autophagy process, the autophagy-related (Atg) genes were discovered in the 1990s. A large body of evidence now indicates that these play a very important role in diseases as neurodegenerative diseases, infectious diseases, and cancers.
The scientists at Horizon Discovery have recognized the importance of having precise tools to study the role of genes and proteins in autophagy. To this end we have developed a panel of cell lines where individual autophagy-related (Atg) genes have been switched off or ”knocked out”, using CRISPR CAS9 gene editing technology. This model system is based on our proprietary cell line the HAP1, and allows the gene edited cell line to be directly compared to the unedited parental cell line. See below, for examples of how our knockout cell line models can help elucidate the genetic components of the autophagy pathway.
Autophagy Response in Hap1 cells
Wild type Hap1 cells were either untreated (Panel A) or stimulated with Bafilomycin A and Torin (Panel B) for 3 hours prior to staining for the autophagy marker LC3. Image analysis using CellProfilerTM software identified autophagosomes (segmented in red in Panels C and D). Quantification of the number of autophagosomes per cell area shows a robust response to stimulation (E), indicating that the Hap1 cells have a functioning autophagic stress response. Images were acquired by the Ketteler Group at the Translational Research Resource Centre at University College London
Atg5 Knockout cells cannot produce autophagosomes
Wild type Hap1 cells (Panel A) or Atg5 KO Hap1 cells (Panel B) were stimulated with Bafilomycin A and Torin (Panel B) for 3 hours prior to staining for the autophagy marker LC3. Image analysis using CellProfilerTM software identified autophagosomes (segmented in red in Panels C and D). Quantification of the number of autophagosomes per cell area shows that the Atg5 KO cells were unable to produce autophagosomes (E).I mages were acquired by the Ketteler Group at the Translational Research Resource Centre at University College London
These examples demonstrate the functionality of Hap1 knockout cells for investigating autophagic responses. The panel of available autophagy cell lines can be a valuable tool in analysing the mechanism of action of a gene or compound involved in autophagic regulation.
For more information on this research area, see:
Choosing Your Ideal Cell Line
Our cell line range is constantly growing. For our most up-to-date list of ready-made cell lines and details of custom made engineering projects, you can browse our cell line range via the following link.
We are so confident you'll be happy with our catalog cell lines, for a limited time, register your interest and we'll even provide a Money Back Guarantee.
For offline access, many of our popular cell lines that are usually available for immediate delivery are listed in our downloadable our catalog - your gene edited cell line may already have been created!