Neural Regen Res. 2022 Oct 10
Di Xu, Li-Ting Zhong, Hai-Yang Cheng, Zeng-Qiang Wang, Xiong-Min Chen, Ai-Ying Feng, Wei-Yi Chen, Gong Chen, and Ying Xu
Products used in the paper Details Operation
AAV vector packaging Viruses were constructed by OBiO Technology (Shanghai, China) or PackGene Biotech (Guangzhou, China). They included AAV7m8 GFAP::GFP-P2A-ND1 (and the control AAV7m8 GFAP::GFP) and AAV9 GFAP104::ND1-P2A-GFP (and the control AAV9 GFAP104::GFP). Request Quote

Research Field: retinal neurons

Keywords: amacrine cell, ganglion cell, horizontal cell, in vivo reprogramming, Müller cell, NeuroD1, photoreceptor, regeneration, retina, retinal degeneration

AAV Serotype: AAV7m8, AAV9

Dose: The titers used in the experiment were 3 × 10^12 GC/mL for AAV7m8 and 1 × 10^12 GC/mL for AAV9, except for the titer test where 10^11, 10^12, and 10^13 GC/mL were used for AAV9.

Routes of Administration: A glass slide with a diameter of 3 mm was placed on the cornea of the mouse so that the fundus could be clearly seen. After puncturing a hole in the corneal limbus with a sharp 30-gauge needle, 0.8–1.2 μL virus was injected into the vitreous (1.2 μL) or subretinal space (0.8 μL) with a 34-gauge blunt-end needle

Targeted organ: eye

Animal or cell line strain: Wild-type (C57BL/6J) male mice


The onset of retinal degenerative disease is often associated with neuronal loss. Therefore, how to regenerate new neurons to restore vision is an important issue. NeuroD1 is a neural transcription factor with the ability to reprogram brain astrocytes into neurons in vivo. Here, we demonstrate that in adult mice, NeuroD1 can reprogram Müller cells, the principal glial cell type in the retina, to become retinal neurons. Most strikingly, ectopic expression of NeuroD1 using two different viral vectors converted Müller cells into different cell types. Specifically, AAV7m8 GFAP681::GFP-ND1 converted Müller cells into inner retinal neurons, including amacrine cells and ganglion cells. In contrast, AAV9 GFAP104::ND1-GFP converted Müller cells into outer retinal neurons such as photoreceptors and horizontal cells, with higher conversion efficiency. Furthermore, we demonstrate that Müller cell conversion induced by AAV9 GFAP104::ND1-GFP displayed clear dose- and time-dependence. These results indicate that Müller cells in adult mice are highly plastic and can be reprogrammed into various subtypes of retinal neurons.

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