Amyloid Precursor Protein and Alzheimer’s

Emma Neale

What is Alzheimer’s?

Link to APP Cascade and Alzheimer’s? Amyloid Precursor Protein (APP) gene is associated with Alzheimer’s

Biological Molecular Cellular

Kunitz

What happens when APP is misregulated?

What happens when APP is misregulated?

Link to APP Cascade and Alzheimer’s? How well conserved is APP? 100%

99%

87%

97%

69%

97%

65%

91%

96%

85%

24%

23%

Kunitz

How related are humans and zebrafish?

Zebrafish APP Alzheimer’s Model

Reward

Zebrafish APP Alzheimer’s model

GAP: Does the APP promoter regulate transcription?

Hypothesis

APPN APPE2Promoter

-3830 -1023 -534 -369-479

Kunitz

GAP: Does the APP promoter regulate transcription?

Hypothesis

APPN APPE2Promoter

-3830 -1023 -534 -369-479

Kunitz

GAP: Does the APP promoter regulate transcription?

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?

What is the primary goal? Understand the role of the

promoter in APP transcription

SequenceMutagenesis ChemicalScreen PhosphorylaHonSites

Aim 1: CRISPR/CAS9

Aim 2: Chemical Genetics

Aim 3: Phosphorylation

SequenceMutagenesis ChemicalScreen PhosphorylaHonSites

APPN APPE2Promoter

-3830 -1023 -534 -369-479

AIM 1: What part of the APP promoter regulates transcription?

Kunitz

APPN APPE2Promoter

-3830 -1023 -534 -369-479

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AIM 1: Use CRISPR to characterize the promoter

SequenceMutagenesis ChemicalScreen PhosphorylaHonSites

Kunitz

AIM 1: Quantify APP transcription rate Hypothesis: mutation will increase APP regulation

WT

X X

X X X

M1

M2

M3

M4

M5

SequenceMutagenesis ChemicalScreen PhosphorylaHonSites

WT Mutant

APP

AIM 2: AD drugs’ affect on Amyloid Beta?

SequenceMutagenesis ChemicalScreen PhosphorylaHonSites

AIM 2: Does the APP promoter affect solubility? Methionine Sulfoxide

Florbetaben Flutemetamol

SequenceMutagenesis ChemicalScreen PhosphorylaHonSites

SequenceMutagenesis ChemicalScreen PhosphorylaHonSites

WT Mutant

AB

AIM 2: Can AD drugs solubilize mutant protein? Hypothesis: Promoter Mutants are soluble

AIM 3: CRISPR’s affect on APP phosphorylation?

SequenceMutagenesis ChemicalScreen PhosphorylaHonSites

Kunitz

ProteaseInhibitor

SequenceMutagenesis ChemicalScreen PhosphorylaHonSites

VS X

?

AIM 3: Does CRISPR affect APP phosphorylation? Hypothesis: Phosphorylation increases proportionally

SequenceMutagenesis ChemicalScreen PhosphorylaHonSites

VS X

AIM 3: Does CRISPR affect APP phosphorylation? Hypothesis: Phosphorylation increases proportionally

Future Directions

Rescue?

Understanding regulation, understanding Alzheimer’s

References https://www.forbes.com/forbes/welcome/?toURL=https://www.forbes.com/sites/alicegwalton/2015/02/09/7-ways-meditation-can-actually-change-the-brain/&refURL=https://www.google.com/&referrer=https://www.google.com/

http://blogs-images.forbes.com/work-in-progress/files/2012/11/brain-puzzle-295x300.jpg?width=960

https://medium.com/enc-3310-spring-2016/the-memory-keepers-understanding-the-communication-break-down-in-alzheimer-s-disease-4c1bdaa7d80d

http://www.abcam.com/amyloid-precursor-protein-antibody-ab15272.html#description_images_2

https://pdb101.rcsb.org/motm/79

http://www.techtimes.com/articles/161325/20160527/new-alzheimers-disease-theory-infections-may-trigger-build-up-of-amyloid-plaques-in-the-brain.htm

http://www.sigmaaldrich.com/life-science/cell-biology/learning-center/bace1-assay-kit.html

http://cordantsolutions.com/president-obama-speaks-on-prescription-drug-abuse/

https://clipartfest.com/categories/view/b08bf1cc755da589799109415f66af287d9310f5/neuron-clip-art.html

http://www.memeron.com/alzheimers.html

http://salarian135.persiangig.com/

http://www.marshfieldresearch.org/chg/pmrp

Questions?

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