Genome-Wide Identification of Cell Type Specific Transcription Factor

Targets During Drosophilia Embyrogenesis

Jessica Osterhout

How are the same transcription factors used multiple times throughout development to specify dramatically different cell fates?

Do they bind and regulate the same target genes in all cases or do they regulates distinct targets in each cell type?

Transcription factors to be tested:

Eve Hb

Chromatin Immunoprecipitaiton followed by sequencing (ChIP-Seq)

FLAG

Hb

Composite

HA

Hb

Composite

Hb Antibody Stains

Blastoderm: stage 5

FLAG Hb Composite

Stage 16

Hb Western Blots

Yw Hb

Stained with Tub

Stained with FLAG

FLAG HA HA

Eve Antibody Stains

Eve Western Blot

To make sure tag does not disrupt Hb protein function: Recombined with two known null alleles to see if tagged Hb rescues.

Future Direction: ChIP-Seq with HA or FLAG at

each stage of development

Acknowledgements and Thanks:

SPUR program

Mike Miller

Chris Doe

Khoa Tran

Bowerman Lab

Prohooda Lab