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Controlling Bacteria With Light

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Presentation on theme: "Controlling Bacteria With Light"— Presentation transcript:

1 Controlling Bacteria With Light
By: Jesse Hinshaw Special Thanks to INBRE, Mark Gomelsky, and Stacey Fomicheva

2 Summary What is optogenetics? Why use NIR-light? What is new
What has been done Current State/Results Conclusion/Continued screening

3 What is Optogenetics? Light Action
Optogenetics is the combination of genetics and optics to control well- defined events within specific cells of living tissue. Light Action

4 Why NIR (Near Infrared) light?
Benign Deeply penetrates mammalian tissue High precision: activation when and where you need it. nm

5 Why Guanylate Cyclase? Guanylate cyclase produce cGMP
cGMP is almost undetectable by pathogen NOT present in most pathogens Cyclase Domain

6 Why Should You Care? Host-Pathogen Studies Mammalian regulation
Heartbeat regulation Memory Replace Viagra

7 How do we know they work? Phytochrome GTP cGMP RNA Polymerase
Guanylate Cyclase Phytochrome GTP cGMP RNA Polymerase Transcription Activator LacZ-Enzyme LacZ Blue Pigment

8 In a perfect world…… Blue colonies White colonies

9 Does the cyclase work? crpA & IlgC3 crpG & IlgC3

10 Random Mutagenesis Mutagenic PCR Ligation Transformation Screen
Harvest Biomass Sequence

11 Almost had it!

12 Challenges Trouble inducing correct number of mutations
Recent sequencing results suggest success: 5-7/Kb Ligation is sometimes difficult Chances of finding correct mutant are small 10^5 Transformants 10^5 Transformants Restreak, collect plasmid, sequence, repeat Harvest Plasmids and transform into crpG cells.

13 Project Future Random mutagenesis of point mutation construct
Continued optimization Move to mice for in vivo analysis Focus on heart regulation or impotence

14 Thanks For the Support! EPSCoR/INBRE Mark Gomelsky Anastasia Fomicheva Other Gomelsky Lab Members

15 Finally, Questions?


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