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Engineer “ArgGlyAsp” (RGD)

Josh Powell virus research

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Page 1: Josh Powell virus research

Engineer  “Arg-­‐Gly-­‐Asp”  (RGD)  

Page 2: Josh Powell virus research

RGD:    stem-­‐cell    

adhesion  epitope  also  target  cancer  

cells  that  up-­‐regulate  integrin  

TYMV  with  RGD  subsDtuDon  at  residue  44  

Page 3: Josh Powell virus research

Wild-­‐type  

N-­‐term:  Mutant  

pH-­‐dependent  enhanced  stability  Mutate  the  internal  scaffold  N-­‐terminus  of  CP    

Page 4: Josh Powell virus research

Robust  TYMV  virus-­‐like  parDcles  in  E.  coli  

 In  vitro  reassembly  from  a  denatured  

state  

Page 5: Josh Powell virus research

AMer  many  months  crystal  structure  data  for  TYMV  and  

mutants!    

Page 6: Josh Powell virus research

TYMV  can  form  a  molecular  “container”  aMer  release  of  RNA,  nano-­‐inspired  applicaDons  with  a  protein  shell  with  a  small  “hole”  

AMer  freeze-­‐thaw  

before  

Page 7: Josh Powell virus research

TYMV  dual-­‐subgenomic  RNA  for  expression  of  GFP  and  GUS.    Future  downstream  use  for  vaccine  development  

Page 8: Josh Powell virus research

Biophysical  studies  of  TYMV  CharacterizaDon  empty,  virion  and  intermediate  parDcles  

Northerns,  Westerns,  gradient  centrifugaDon,  etc.  

Page 9: Josh Powell virus research

Previous  Work  (masters):    Signaling  of  a  Cytomegalovirus  (CMV)  chemokine  receptor  US28:  plenty  of  cloning,  Dssue  culture,  immuno-­‐precipitaDon/fluorescence  and  construcDng  CMV  knockout  mutants  

US28  chemokine  

Page 10: Josh Powell virus research

Recent  TYMV  arDcle    &  (cover  image)    Jan  2011,  more  publicaDons  to  come!