Skip to main content
. 2019 Oct 23;15:320–332. doi: 10.1016/j.omtm.2019.10.007

Figure 1.

Figure 1

iTransduce Library for Selection of Novel AAV Capsids Capable of Efficient Transgene Expression in Target Tissue

(A) Two-component system of the library construct. (1) Cre recombinase is driven by a minimal chicken β-actin (CBA) promoter. (2) p41 promoter-driven AAV9 capsid with random heptamer peptide is inserted between amino acids 588 and 589, cloned downstream of the Cre cassette. (B) Selection strategy. (Bi) The iTransduce library comprised of different peptide inserts expressed on the capsid (represented by different colors) is injected intravenously (i.v.) into an Ai9 transgenic mouse with a loxP-flanked STOP cassette upsteam of the tdTomato reporter gene, inserted into the Gt(ROSA)26Sor locus. AAV capsids able to enter the cell of interest but that do not functionally transduce the cell (no Cre expression) do not turn on tdTomato expression. Capsids that can mediate functional transduction (express Cre) will turn on tdTomato expression. (Bii) Cells are isolated from the organ of interest (e.g., brain), and transduced cells are sorted for tdTomato expression and optionally cell markers. (Biii) Capsid DNA is PCR amplified from the sorted cells, cloned back to the library vector, and repackaged for another round of selection. DNA sequencing analysis is utilized after each round to monitor the selection process.