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85 INTRODUCTION 87 MATERIALS AND METHODS 90 Mosquito Maintenance and Injection Protocol 90 Establishing Brugia Infection 90 Gene Selection & dsRNA Generation 91 Relative Quantitative RT-PCR 93 Quantitative RT-qPCR 95 Phenotype Analysis 98 Statistical Analysis 99 RESULTS 100 Target Genes were Expressed During Mosquito Life Cycle Stages 100 In Squito Suppression of Target Genes was Potent and Specific 103 Brugia Gene Suppression Result in Aberrant Motility 110 Effect of dsRNA Exposure on Normal Knotting Behavior of L3 B. malayi 113 Worm Distribution is Significantly Altered with RNAi Suppression 116 Effect of B. malayi Target Gene Suppression on Worm Survival 120 Effect of B. malayi Target Gene Suppression on Mosquito Survival 120 DISCUSSION 122 The Robustness of the “In Squito” RNAi Protocol 122 Bm-tub-1 122 Bm-GluCl-α3A 123

The development and utilization of an in vivo RNA interference protocol to elucidate gene functions and identify potential drug targets in the filarial nematode Brugia malayi