ZFIN Image: Tucker et al., 2021, Fig. 1

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Fig. 1

Design and generation of MAK gene transfer constructs.

A Schematic depicting the four AAV constructs generated in this study. Constructs were designed to deliver either the human canonical (MAK^CI - lacks exon 12, constructs 1 and 3) or retina-specific MAK isoform (MAK^RI - contains exon 12, constructs 2 and 4) under control of either the constitutively active CMV or EF1α promoter. B Representative image of cultured murine JK1 cells (immortalized SMA + , CD34 + testicular stromal cells, which lack endogenous expression of human MAK). Scale bar 100 μm. C, D rt-PCR analysis performed on RNA isolated from JK1 cells transduced with either AAV5-CMV-MAK^CI (Construct 1), AAV5-CMV-MAK^RI (Construct 2), AAV5-EF1α-MAK^CI (Construct 3), or AAV5-EF1α-MAK^RI (Construct 4). Each construct is capable of driving robust expression of exon-9-containing human MAK transcript (C). Unlike the MAK^CI constructs, both AAV5-CMV-MAK^RI and AAV5-EF1α-MAK^RI constructs are capable of driving expression of the exon-12-containing retina-specific MAK transcript (D). Arrows denote rt-PCR primer positions.

Acknowledgments

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