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  Indian J Med Microbiol
 

Figure 5: Schema of replication-deficient herpes simplex virus (HSV) vectors. (a) The vector backbone vLG is generated from the HSV KOS strain in which BAC sequences are inserted into the TK locus (UL23). Deletions are introduced in the internal repeat region and the genes encoding immediate early proteins ICP4 and ICP27, rendering the vector replication-defective. The Gateway destination cassette is inserted into the remaining latency locus, replacing the latency promoter elements while maintaining a wildtype copy of ICP0 and the surrounding CTCF chromatin boundary elements. (b) To generate afferent neuronal subtype-targeted expression vectors, transgenes are recombined into the vector backbone via the Gateway cassette. Individual promoter sequences (1284 bp TRPV1 promoter [TRPV1p], 932 bp CGRP promoter [CGRPp], 553 bp CMV promoter [CMVp], and 970 bp NF200 promoter [NF200p]) incorporating a Kozak consensus translation initiation sequence are inserted into the pENTR1A transfer vector, which contains attL sites for site-directed recombination with the attR-containing vector backbone. These Gateway plasmids are then recombined into the vLG vector backbone to generate the experimental vectors. (c) Schema of vectors. Replication-deficient HSV vectors in combination with wild type (upper) and the double mutant (G2M) (lower) α1 glycine receptor subunit gene. The study (Saito, et al., 2019[107]) utilized a chemogenetic approach using a G2M receptor with increased sensitivity to ivermectin, in combination with replication-defective HSV vector-mediated gene delivery methods. HSV: Herpes simplex virus

Figure 5: Schema of replication-deficient herpes simplex virus (HSV) vectors. (a) The vector backbone vLG is generated from the HSV KOS strain in which BAC sequences are inserted into the TK locus (UL23). Deletions are introduced in the internal repeat region and the genes encoding immediate early proteins ICP4 and ICP27, rendering the vector replication-defective. The Gateway destination cassette is inserted into the remaining latency locus, replacing the latency promoter elements while maintaining a wildtype copy of ICP0 and the surrounding CTCF chromatin boundary elements. (b) To generate afferent neuronal subtype-targeted expression vectors, transgenes are recombined into the vector backbone via the Gateway cassette. Individual promoter sequences (1284 bp TRPV1 promoter [TRPV1p], 932 bp CGRP promoter [CGRPp], 553 bp CMV promoter [CMVp], and 970 bp NF200 promoter [NF200p]) incorporating a Kozak consensus translation initiation sequence are inserted into the pENTR1A transfer vector, which contains attL sites for site-directed recombination with the attR-containing vector backbone. These Gateway plasmids are then recombined into the vLG vector backbone to generate the experimental vectors. (c) Schema of vectors. Replication-deficient HSV vectors in combination with wild type (upper) and the double mutant (G2M) (lower) α1 glycine receptor subunit gene. The study (Saito, <i>et al</i>., 2019<sup>[107]</sup>) utilized a chemogenetic approach using a G2M receptor with increased sensitivity to ivermectin, in combination with replication-defective HSV vector-mediated gene delivery methods. HSV: Herpes simplex virus