Development of CRISPR/Cas9-based Novel Vaccines against Poultry Viruses

Vaccines remain the primary means of disease prevention throughimmunisationschemes in the poultry sector. Novel approaches in vaccine development, such as reverse genetic systems and genome editing technologies (i.e. CRISPR/Cas9), are currently being utilized to overcome challenges in establishing an immunogenic platform that is safe and capable of inducing long-term immunity. The CRISPR/Cas9 (Clustered regularly interspaced short palindromic repeats associated protein nuclease 9) technology offers an effective, fast and simple novel approach to edit genomes for the development of viral vectors against poultry diseases such as Newcastle disease (ND), avian influenza (AI) and infectious bronchitis (IB). In this study, we demonstrate the application of CRISPR/Cas9-based genome editing to generate recombinant viral vectors that express the F gene of NDV. Validation of gene integration, protein expression, and insert stability was carried out by PCR, immunostaining and Western blot analysis, respectively. This approach offers an efficient platform for the generation of multivalent recombinant vaccines that can provide simultaneous protection against major poultry diseases.