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Our customer‘s goal was to rapidly evaluate a set of AAV constructs in vitro to compare their transduction efficiency and verify the cell types transduced in a neural retina model.
We followed our standardised series of steps to develop a customised solution.
We clearly defined the problem and questions to be answered through in-depth discussion between the customer and our technical experts.
The customer wished to compare the transduction efficiency of a series of AAV vectors with different capsids, promoters and transgenes.
Taking all project specific factors into account, we suggested a two-step approach as being the most appropriate for their situation:
We designed the specific experiments and presented the plan in our Statement of Work.
Our experimental plan involved using fully validated human retinal organoids.
For Step 1, transduce organoids at day 150 of differentiation with AAV vectors and live imaging from 5 to 27 days post-transduction. The experiments aimed to evaluate the transduction efficiency of the various AAV constructs. We also checked that organoid viability was not negatively affected by AAV transduction.
For Step 2 we planned to carry out the following imaging analysis,:
Our scientists carried out the agreed experiments within 3 months providing regular updates to the customer.
The study included the experimental phase, data processing, data analysis and presentation of a data summary.
Example dataset
Imaging comparison of a series of AAV construct with different promotors, reporter genes and capsids.
We delivered a detailed report with reliable dataset which was shared digitally and discussed over a call upon request.
The study clearly showed :
This study expanded the options of AAV capsid and transgene to be used for preclinical testing of gene therapy to include CAG quad mutant variants and GRK1 into AAV vectors
We delivered a robust dataset which clearly showed that human retinal organoids are a robust in vitro model for rapid screening of AAV constructs. Our analysis gave confidence to our customer to select the most effective AAV construct.
Get rapid and comprehensive compound efficacy & safety evaluation with increased sensitivity for confident decision-making. Enquire today about how this assay can bring you more insights!