Apoptosis/Killing Tools

Abnormalities in programmed cell death (PCD) signaling cascades, including apoptosis, necrosis, pyroptosis, ferroptosis, autophagy-related cell death, and non-programmed necrosis, can be observed in the pathogenesis of various neurological diseases. These cell deaths can occur in reaction to numerous types of cellular stress (caused by intracellular or extracellular stimuli) and inflammation. Aberrant activation of PCD pathways is a common feature of neurodegenerative diseases, such as amyotrophic lateral sclerosis (ALS), Alzheimer's disease, Parkinson's disease, and Huntington's disease, leading to unexpected losses of neuronal cells and functions.

Our apoptosis/killing tools can help ablate target cells to study their specific functions. For in vivo model applications, apoptosis/killing tools can also simulate the abnormal activation of PCD pathways in neurodegenerative diseases, helping researchers to elucidate the underlying mechanisms of the disease and identify potential treatments for the disease.

Applications of Apoptosis/Killing Tools

• taCasp3-TEVp induces apoptosis

Casp3 protein is a cell apoptosis effector molecule. During cell apoptosis, molecules such as caspase-10 will cleave Casp3 between positions 175 and 176, activating the activity of Casp3. taCasp3 has the recognition site of TEVp inserted at this position, a specific serine protease derived from the Tobacco Etch Virus. When expressed in cells, TEVp is first expressed and then activates Casp3, triggering a cascade reaction leading to apoptosis.

• Diphtheria toxin A (DTA) induces cell death

DTA is the fragment of diphtheria toxin, which can inhibit protein synthesis and cell death. DTA functional elements can be used to selectively eliminate specific cells to study the unique functions of these cells and are also widely used to model neurological diseases.

Case Study

In vivo, virally encoded DTA or tBid are only partially successful at ablating neurons. Using genetically modified caspase 3, activating this caspase triggers apoptosis, which ablates adult neurons in vivo. AAV-flex-taCasp3-TEVp regulates the expression of pro-taCasp3 and TEVp in a Cre-dependent way.

Fig.1 taCasp3-TEV ablates neurons in a Cre-dependent manner. A-B) Workflow of the genetic strategy for conditional ablation of neurons. C-D) One week after Cre:EGFP+ HEK293T cells were infected with AAV-flex-taCasp3-TEVp, there was cell death. E-F) Neuronal ablation in mice with AAV-flex-taCasp3-TEVp injection.¹

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Reference

1. Yang, Cindy F., et al. "Sexually dimorphic neurons in the ventromedial hypothalamus govern mating in both sexes and aggression in males." Cell 153.4 (2013): 896-909.