SCREEN FOR DOMINANT BEHAVIORAL MUTATIONS CAUSED BY GENOMIC INSERTION OF P-ELEMENT TRANSPOSONS IN DROSOPHILA: AN EXAMINATION OF THE INTEGRATION OF VIRAL VECTOR SEQUENCES

Abstract

Here we report the development of a high-throughput screen to assess dominant mutation rates caused by P-element transposition within the Drosophila genome that is suitable for assessing the undesirable effects of integrating foreign regulatory sequences (viral cargo) into a host genome. Three different behavioral paradigms were used: sensitivity to mechanical stress, response to heat stress, and ability to fly. The results, from our screen of 35,000 flies, indicate that mutations caused by the random insertion of transposons in Drosophila are more effective at disrupting flight than stress sensitivity. This approach was used to ascertain the frequency of deleterious dominant mutations caused by viral vectors utilized in gene therapy.

Keywords:

• Adeno Associated Virus (AAV)
• Complex behaviors
• Cytomegalovirus (CMV)
• Gene therapy
• Mutagenesis
• Simian Virus-40 (SV-40)
• Stress sensitivity