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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