About the Ac/Ds screen

We are using the maize Ac/Ds transposon system to carry out an insertional mutagenesis screen in zebrafish. The Ac/Ds transposon system yields high rate of germ-line transmission in zebrafish (Emelyanov et al., 2006). We have generated around 300 Ds transgenic insertion lines, and find that transgenic founders, on average, harbor 3 inserts. Expression of gene trap fluorescent reporter in progeny of injected founders was found in multiple cell types during embryonic and larval stages. F1 progeny of the Ds insertion lines have been confirmed by PCR, and so far, flanking sequences from ~300 insertion lines have been obtained.

We have performed phenotypic characterization of F2 fish, and have identified insertion mutants with defects in the digestive organs, CNS, heart, etc. Our gene trap insertion screen shows that the maize Ac/Ds system can be successfully to trap genes in zebrafish, and can complement screens using the Tol2 and retroviral systems to achieve saturation.


View construct and sequences used