Establishing genetic transformation for comparative developmental studies in the crustacean Parhyale hawaiensis.
The amphipod crustacean Parhyale hawaiensis has been put forward as an attractive organism for evolutionary developmental comparisons, and considerable effort is being invested in isolating developmental genes and studying their expression patterns in this species. The scope of these studies could be significantly expanded by establishing means for genetic manipulation that would enable direct studies of gene functions to be carried out in this species. Here, we report the use of the Minos transposable element for the genetic transformation of P. hawaiensis. Transformed amphipods can be obtained from approximately 30% of surviving individuals injected with both a Minos element carrying the 3xP3-DsRed fluorescent marker and with mRNA encoding the Minos transposase. Integral copies of the transposon are inserted into the host genome and are stably inherited through successive generations. We have used reporter constructs to identify a muscle-specific regulatory element from Parhyale, demonstrating that this transformation vector can be used to test the activity of cis-regulatory elements in this species. The relatively high efficiency of this transgenic methodology opens new opportunities for the direct study of cis-regulatory elements and gene functions in Parhyale, allowing functional studies to be carried out beyond previously established model systems in insects.
Pavlopoulos
Anastasios
A
Institute of Molecular Biology and Biotechnology, 711 10 Iraklio Crete, Greece. ap448@cam.ac.uk
Averof
Michalis
M
D033304:Amphipoda / Q000235:genetics*; D000818:Animals; D003001:Cloning, Molecular; D017931:DNA Primers; D004251:DNA Transposable Elements / Q000235:genetics; D015509:Developmental Biology / Q000379:methods*; D018014:Gene Transfer Techniques; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics; D008164:Luminescent Proteins / Q000235:genetics; D008845:Microinjections; D023421:Models, Animal; D010957:Plasmids / Q000235:genetics; D014170:Transformation, Genetic / Q000235:genetics*; D019895:Transposases / Q000235:genetics
D017931:DNA Primers; D004251:DNA Transposable Elements; D018840:HSP70 Heat-Shock Proteins; D008164:Luminescent Proteins; C413663:fluorescent protein 583; C473499:Minos transposase; D019895:Transposases