Differential Display PCR - what is it?

Basavaraju Shankarappa bsh at MED.PITT.EDU
Tue Apr 12 11:13:14 EST 1994


> Hi folks,
> What is the above?
> Thanks 
> Duncan
> -----------------------------------------------------------------------------
> Duncan Clark                        | Internet:    duncan at genesys.demon.co.uk
> GeneSys Ltd.                        | Compuserve:  100015.1406 at compuserve.com
> -----------------------------------------------------------------------------

Hi Duncan:	
	
In my opinion this happens to be one of the technology of the future
that will be used exclusively in the future to identify unique
transcripts when the effect of a particular treatment is to be
studied.  In this method selected population of cDNAs are made using
polyT-CA (for eg.,) followed by amplification with this primer and
an upstream arbitraty primer.  This will generate a unique fingerprint
of the message dependent on cell type, type of polyT used and the
type or arbitrary primer used.  By comparing the standard and the
treated group of cells, you can identify the presence of the isolate
the sequences corresponding to the messaged that are uniquely present
in a cancerous cell as opposed to its normal cell counterpart, identify
the messages that are expressed following infection with a pathogen
or with a drug and as far as I am concerned the list can go on pretty
far.
	Following are some of the references based on Liang and Pardee's
work.  Incidentally, McClelland also reported similar but not same
technique about the same time in NAR (Welsh et al., Arbitrarily primer
PCR fingerprinting of RNA. NAR 20:4965).  But their technique was not
specific to mRNA whereas Liang and Pardees technique is, I believe,
preferential but not exclusive to mRNA in view of the use of
polyT-NN for making cDNA.  
  
1.  Bauer D.  Muller H.  Reich J.  Riedel H.  Ahrenkiel V.
    Warthoe P.  Strauss M.
  Title
    Identification of differentially expressed mRNA species
    by an improved display technique (DDRT-PCR).
  Source
    Nucleic Acids Research.  21(18):4272-80, 1993 Sep 11.
2.    Zhang L.  Medina D.
   Title
   Gene expression screening for specific genes associated
   with mouse mammary tumor development.
  Source
    Molecular Carcinogenesis.  8(2):123-6, 1993.
3.  Authors
    Sager R.  Anisowicz A.  Neveu M.  Liang P.  Sotiropoulou G.
  Title
    Identification by differential display of alpha 6
    integrin as a candidate tumor suppressor gene.
  Source
    FASEB Journal.  7(10):964-70, 1993 Jul.
4.  Liang P.  Averboukh L.  Pardee AB.
  Title
    Distribution and cloning of eukaryotic mRNAs by means of
    differential display: refinements and optimization.
  Source
    Nucleic Acids Research.  21(14):3269-75, 1993 Jul 11.
5.  Authors
    Liang P.  Pardee AB.
  Title
    Differential display of eukaryotic messenger RNA by
    means of the polymerase chain reaction.
  Source
    Science.  257(5072):967-71, 1992 Aug 14.
6.  Authors
    Liang P.  Averboukh L.  Keyomarsi K.  Sager R.  Pardee AB.
  Title
    Differential display and cloning of messenger RNAs from
    human breast cancer versus mammary epithelial cells.
  Source
    Cancer Research.  52(24):6966-8, 1992 Dec 15.

Raj Shankarappa 
bsh at med.pitt.edu




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