A computer-implemented method for generating tripwire files may include (1) generating an initial tripwire file according to an initial tripwire generation calculation, the initial tripwire file configured such that modification of the initial tripwire file triggers investigation of a security breach, (2) generating a subsequent tripwire file according to a subsequent tripwire generation calculation, the subsequent tripwire generation calculation differing from the initial tripwire generation calculation along at least one dimension, (3) receiving automated feedback that indicates whether at least one of the initial tripwire file and the subsequent tripwire file failed to enable detection of a security threat, and (4) adjusting automatic generation of a third tripwire file based on the automated feedback indicating whether at least one of the initial tripwire file and the subsequent tripwire file failed to detect the security threat. Various other methods, systems, and computer-readable media are also disclosed.