The w+mW.hs marker from the progenitor TI{TI}tauKO is removed via P1\cre, followed by insertion of sequence at the attP site within TI{TI}tauKO. The inserted sequence consists of full-length pseudo-de-acetylated Hsap\MAPT 2N4R cDNA (contains mutation to arginine of 4 acetylation sites: K163, K280, K281 and K369), along with a loxP-flanked w+mW.hs marker and an ampicillin resistance gene.