Fig. 2

Construction of FOXO3 donor vector. A fragment of FOXO3 (Arm 1) was inserted into the pcDNA3 vector proximal to the DraIII restriction site by using Gibson assembly. This intermediate vector was called FOXO3 Arm1 and was utilized to make the final FOXO3 donor vector. Another fragment of FOXO3 (Arm 2) was inserted into the FOXO3 Arm1 vector (by Gibson assembly) to obtain the final FOXO3 donor vector. This vector was confirmed by Sanger sequencing and employed in CRISPR Cas9 mutagenesis reactions