AAV Production for Autophagy Flux Detection

Autophagy is a catabolic mechanism which sequesters cytoplasmic components for lysosomal-mediated degradation, including protein inclusions and damaged organelles. Autophagy has an important role in diverse physiological processes such as adaptation to nutrient starvation, cell death, embryonic development, energy metabolism, and antigen presentation, among other events. Moreover, autophagy has been highlighted as a crucial component of pathological processes such as cancer, hypoxia, pathogen infection, and neurodegeneration. Thus, the estimation of the rate of autophagy in a quantitative manner, especially in in vivo models, has become a vital need in the field.

One of the important steps in the regulation of autophagy is the conjugation of microtubule-associated protein 1 light chain 3 (LC3) to phosphatidylethanolamine to form microtubule-associated protein 1 light chain 3 lipidated form (LC3-II). Lipidated LC3 binds to the expanding phagophore and remains associated with autophagosomes even after fusion with lysosomes. Then, LC3-II can be either delipidated and recycled or degraded by hydrolytic enzymes at the lysosome. Monitoring LC3-II conversion, LC3 distribution or its flux through the autophagy pathway are the gold standards for measuring autophagy activity and are widely used in the field. Recombinant adeno-associated vector (AAV) vectors can efficiently transfect various cell types, including dividing and quiescent cells, and induce persistent gene expression in vivo without integrating into host genome and causing any disease. These features make AAV an attractive candidate in the application of gene delivery for gene therapy and human disease model establishment. Besides, adeno-associated virus can choose from among different serotypes relying on the affinity to tissues and organs, which is the most effective tool for in vivo detection of autophagy flux.

Schematic representation of the fluorescent reporter viral delivery strategy and the autophagy flux analysis.

Figure 1. Schematic representation of the fluorescent reporter viral delivery strategy and the autophagy flux analysis.

Our AAV-LC3 Production Service for Autophagy Flux Detection

QVirus™ Platform has launched series of adenovirus packaging service of autophagy-related biosensors, in which GFP and/or RFP tags are fused at the C-termini of the autophagosome marker LC3, allowing to detect the intensity of autophagy flux in real-time with more accuracy, intuitiveness and clarity.

Features

➢ Real time and quantitative in vivo autophagy flux detection.
➢ High resolution, more sensitivity and accuracy, than traditional approaches.
➢ Broad range of host and specificity targeting.
➢ Safety and low immunogenicity.
➢ Stable physical properties.

Applications

  • Measurement of autophagy flux in cultured cells
  • Monitoring autophagy in animals
  • Drug discovery study

QVirus™ Platform provides AAV production services to study the different stages of autophagy flux, making the autophagy study much easier. If you have any special requirements, please feel free to contact us.

References
1. Castillo K, et al. Measurement of autophagy flux in the nervous system in vivo. Autophagy, 2014, 4(4):e917-714.
2. Matus S, et al. A new method to measure autophagy flux in the nervous system. Autophagy, 2014, 10(4):710-714.

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