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Licensing opportunity DI 05/40

Recovery of APOBEC3-edited human immunodeficiency virus A G hypermutants by differential DNA denaturation PCR

Inventors: S. Wain-Hobson et al.

Description of invention: 

DNA complementarity is expressed by way of three hydrogen bonds for a G:C base pair and two for A:T. As a result, careful control of the denaturation temperature of PCR allows selective amplification of AT-rich alleles. Inosine (I) hydrogen bonds to cytosine by two hydrogen bonds while diaminopurine (D) forms three hydrogen bonds with thymine. By substituting dATP and dGTP by dDTP and dITP in a PCR reaction, DNA is obtained in which the natural hydrogen bonding rule is now inverted. When PCR is so performed at limiting denaturation temperatures GC-rich alleles resulting from editing by dsRNA dependent host cell adenosine deaminases can be selectively recovered from both measles and Rift Valley fever virus cultures. The method can be extrapolated to the detection of any GC-rich allele or microbial strain.

 Specific advantages

 A quick and easy to perform method.

 Potential applications
 The identification of GC rich bacterial or microbial sequences in the presence of other bacteria. The identification of double or multiple infection by various strains if the same microbe that differ in their GC/AT content. The identification of adenosine edited mRNAs. The identification and quantitation of GC rich DNA.
Patent Status:
 US patent application filed on December 30, 2005
 International patent application filed on December 28, 2006

 Recent pertinent publications:
 Interferon-inductible expression of APOBEC3 editing enzymes in human hepatocytes and inhibition of hepatitis B virus replication
Bonvin M, Achermann F, Greeve I, Stroka D, Keogh A, Inderbitzin D, Candinas D, Sommer P, Wain-Hobson S, Vartanian JP, Greeve J.
Hepatology. 2006 Jun;3(6):1364-74. 
 Restriction of Foamy viruses bye APOBEC cytidine deaminases
Delebecque F, Suspene R, Calattini S, Casartelli N, Saib A, Froment A, Wain-Hobson S, Gessain A, Vartanian JP, Schwartz O.
J Virol. 2006 Jan;80(2):605-14.
 Extensive editing of a small fraction of human T-cell leukemia virus type 1 genomes by four APOBEC3 cytidine deaminases
Mahieux R, Suspene R, Delebecque F, Henry M, Schwartz O, Wain-Hobson S, Vartanian JP.
J Gen Virol. 2005 Sep;86(Pt 9):2489-94.

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