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. 2018 Oct 15;3(6):e695. doi: 10.1097/PR9.0000000000000695

Figure 1.

Figure 1.

Construction and administration of HDAd containing DRG-specific motif in the fiber. (A) DNA coding for DRG homing peptide (SPGARAF) was cloned into the BspEI site present in the linker peptide (GGSGGG) between amino acid number 542 and 543 in the HI loop of adenoviral fiber protein. DNA sequence coding for the peptide is shown under the amino acid sequence. Underline shows the BspEI site. A, alanine; F, phenylalanine; G, glycine; P, proline; R, arginine; S, serine; T, threonine. (B) Schematic presentation for the construction of helper virus with a chimeric fiber containing DRG homing peptide (HV-DRG) and HDAd expressing the glutamic acid decarboxylase (GAD) 67 gene (HDAd-GAD67). ITR, inverted terminal repeat; Ψ, packaging signal; EF-1, elongation factor 1 promoter. (C) Schematic presentation of the final step in generating HDAd expressing GAD67 with DRG homing peptide (HDAd-DRG-GAD67). HDAd-DRG-GAD67 was constructed in 293Cre cells by coinfecting HDAd-GAD67 and fiber-modified HV. Helper virus genome is not packaged into HDAd-DRG-GAD67 because the packaging signal (Ψ) of HV is excised by Cre in 293Cre cells. Open circle indicates wild-type fiber; black circle indicates DRG fiber. (D) Schematic presentation of neuropathic pain model of mouse induced by spinal nerve transection (SNT). Lumbar (L)4 SNT induces mechanical allodynia in the ipsilateral hind paw. We administrated therapeutic vectors into the L4 spinal nerve immediately after SNT. Right panel shows an enlargement of the transection site. DRG, dorsal root ganglion; HDAd, helper-dependent adenovirus; HV, helper virus.