Although many in vitro studies have helped us understand how Dicer-2 is able to discriminate between different dsRNA substrate termini, much less is known about how this translates to the in vivo recognition of viral dsRNA. Indeed, Dicer-2 associates with several dsRNA-binding proteins (dsRBPs), which can modify its specificity for a substrate, however it remains unknown how Dicer-2 is able to recognize the protected termini of viral dsRNAs. Therefore, in order to study how the ribonucleoprotein network of Dicer-2 impacts antiviral immunity, we used an IP-MS approach to identify interactants of several fly lines expressing different versions of GFP:Dicer-2. By combining the global analysis of the Dicer-2 interactome with different line-specific analyses, we were able to both obtain a global overview of the partners of Dicer-2 in vivo, and study how this interactome was modulated by different factors such as the infection and/or the presence of different point mutations on the helicase or RNase III domains of GFP:Dicer-2. This allowed the identification of several new Dicer-2 interactants as well as new pro- and antiviral factors that had an impact on DCV infection. In addition, this work provides a resource composed of several candidates, available to the scientific community that can now be investigated further to gain a better understanding of the proteins involved in Dicer-2-mediated antiviral RNAi.