On 3 April 2014, Directive 2014/41/EU regarding the European investigation order in criminal matters was finally approved. This instrument seeks to overcome the present fragmentary regulation on the gathering and transfer of cross-border evidence and improve the efficiency of the international judicial cooperation among the Member States. This article analyses the scope, content, and requirements of the European investigation order in order to make an assessment of this Directive from the point of view of the protection of the fundamental rights envisaged in Article 48 of the European Charter of Fundamental Rights, and, in particular, the right of defence in transnational criminal proceedings.