Clone Repositories at HIP

1
Clone Repositories at HIP
http//plasmid.med.harvard.edu/PLASMID
DF/HCC DNA Resource Core 40,000 clones
PSI-MR 65,000 clones

• FLEXGene
• ORF clones
• Human, yeast Prokaryotes
• Genome-scale collections
• Full-length ORFs
• Fully sequence validated

• Others
• Empty vectors molecular techniques
• Many organisms
• shRNAs, genomic fragments, cDNAs
• Full-length, partial and mutant forms

• PSI Labs
• Expression vectors (various systems)
• Expression clones
• Many organisms
• Full-length, partial and mutant ORFs

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Bench-top Profiling of Individual Immune Response
Protein signal
DNA signal
In situ synthesis
Whole Proteome Array
3
Probing Protein Microarrays
Test sera
Identify which antigens are specific to patient
sera Validation and Discovery in One Step
Replicate microscopic arrays of candidate antigens

• Other Applications of Target Protein Microarrays
• Drug selectivity Which kinases are bound by
  candidate inhibitor?
• Unintended drug targets Which unintended
  proteins bind to drug?
• Enzyme substrate identification Which proteins
  are ubiquitylated?
• Protein-protein interactions whole proteins or
  domains
• Protein mutational analysis Which protein
  domains are key for function?

4
Innate Immune Assay Design
RAW 264.7 Macrophage Cells
TLR

• Macrophage cells have most TLRs
• Pathogen-Associated Molecular Pattern (PAMP)
• LPS
• Flagellin
• Lipoprotein
• Peptidoglycan
• Whole protein extract has extraordinary response
• Goal Find all proteins that induce response

TNFa Detection
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Proteins that Induce Innate Immunity