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Genome-Wide Identification of Cell Type Specific Transcription Factor Targets During Drosophilia Embyrogenesis Jessica Osterhout
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Genome-Wide Identification of Cell Type Specific Transcription Factor Targets During Drosophilia Embyrogenesis Jessica Osterhout.

Dec 13, 2015

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Page 1: Genome-Wide Identification of Cell Type Specific Transcription Factor Targets During Drosophilia Embyrogenesis Jessica Osterhout.

Genome-Wide Identification of Cell Type Specific Transcription Factor

Targets During Drosophilia Embyrogenesis

Jessica Osterhout

Page 2: 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

Page 3: Genome-Wide Identification of Cell Type Specific Transcription Factor Targets During Drosophilia Embyrogenesis Jessica Osterhout.

Chromatin Immunoprecipitaiton followed by sequencing (ChIP-Seq)

Page 4: Genome-Wide Identification of Cell Type Specific Transcription Factor Targets During Drosophilia Embyrogenesis Jessica Osterhout.
Page 5: Genome-Wide Identification of Cell Type Specific Transcription Factor Targets During Drosophilia Embyrogenesis Jessica Osterhout.

FLAG

Hb

Composite

HA

Hb

Composite

Hb Antibody Stains

Blastoderm: stage 5

Page 6: Genome-Wide Identification of Cell Type Specific Transcription Factor Targets During Drosophilia Embyrogenesis Jessica Osterhout.

FLAG Hb Composite

Stage 16

Page 7: Genome-Wide Identification of Cell Type Specific Transcription Factor Targets During Drosophilia Embyrogenesis Jessica Osterhout.

Hb Western Blots

Yw Hb

Stained with Tub

Stained with FLAG

Page 8: Genome-Wide Identification of Cell Type Specific Transcription Factor Targets During Drosophilia Embyrogenesis Jessica Osterhout.

FLAG HA HA

Eve Antibody Stains

Page 9: Genome-Wide Identification of Cell Type Specific Transcription Factor Targets During Drosophilia Embyrogenesis Jessica Osterhout.

Eve Western Blot

Page 10: Genome-Wide Identification of Cell Type Specific Transcription Factor Targets During Drosophilia Embyrogenesis Jessica Osterhout.

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

Page 11: Genome-Wide Identification of Cell Type Specific Transcription Factor Targets During Drosophilia Embyrogenesis Jessica Osterhout.

Future Direction: ChIP-Seq with HA or FLAG at

each stage of development

Page 12: Genome-Wide Identification of Cell Type Specific Transcription Factor Targets During Drosophilia Embyrogenesis Jessica Osterhout.

Acknowledgements and Thanks:

SPUR program

Mike Miller

Chris Doe

Khoa Tran

Bowerman Lab

Prohooda Lab