Gene duplicates and their involvement in development of novel traits in butterflies: a study on engrailed paralogs Tirtha Das Banerjee and Antónia Monteiro Email:
[email protected] (TDB);
[email protected] (AM)
Introduction
Engrailed paralogs in Bicyclus anynana
Duplication of DNA region which contains a gene Mechanism: Unequal crossing over, horizontal gene transfer retro transposition and whole genome duplication
Methods CRISPR-Cas9
In-situ hybridization DDC model of gene duplication
Engrailed/Invected in Bicyclus anynana Cas9
Cas9-sgRNA hybrid
Genotyping
sgRNA
Injections
Rearing
cDNA based verification of en paralogs
Results
In-situ hybridization en2
en1
Posterior embryonic segments
inv
Posterior embryonic segments
Posterior embryonic segments
CRISPR-Cas9
en1
Posterior larval wings Eyespot
Androconia
Veins T7 endonuclease assay
Upper posterior larval wings
Posterior larval wings
Eyespot
Eyespot
Androconia
Veins
Veins
Sequencing
Discussion
… Future Work
• The results indicate that expression in the focal halo, in the posterior larval wings, in the posterior embryonic Perform comparative study with Junonia coenia. Perform CRISPR-Cas9 on en2 and inv and analyze their function. segments, and along the veins are ancestral to all the three copies. • inv lost expression in the lower posterior compartment, while en1 and en2 gained expression in the androconial cells. • These results suggest that the sequences that lead to en/inv expression in eyespots predate the origin of eyespots. Trans-regulatory changes in genes upstream of en/inv might have lead to the co-option of these genes to the eyespot Diane Ramos for initial work and Monteiro Lab members for region. their support.
Acknowledgement
