Jongmans,
Taking your vacation early this year? Haven't heard from ye in a while.
Very interesting description of powerful splicing technique from PNAS and DoubleTwist. Note one of the authors is now at Lexicon.
>>High efficiency mutagenesis, repair, and engineering of chromosomal DNA using single-stranded oligonucleotides
Hilary M. Ellis, Daiguan Yu*, Tina DiTizio, and Donald L. Court
Gene Regulation and Chromosome Biology Laboratory, Division of Basic Sciences, National Cancer Institute at Frederick, P.O. Box B, Frederick, MD 21702
Communicated by Sankar Adhya, National Institutes of Health, Bethesda, MD, April 3, 2001 (received for review February 16, 2001)
Homologous DNA recombination is a fundamental, regenerative process within living organisms. However, in most organisms, homologous recombination is a rare event, requiring a complex set of reactions and extensive homology. We demonstrate in this paper that Beta protein of phage generates recombinants in chromosomal DNA by using synthetic single-stranded DNAs (ssDNA) as short as 30 bases long. This ssDNA recombination can be used to mutagenize or repair the chromosome with efficiencies that generate up to 6% recombinants among treated cells. Mechanistically, it appears that Beta protein, a Rad52-like protein, binds and anneals the ssDNA donor to a complementary single-strand near the DNA replication fork to generate the recombinant. This type of homologous recombination with ssDNA provides new avenues for studying and modifying genomes ranging from bacterial pathogens to eukaryotes. Beta protein and ssDNA may prove generally applicable for repairing DNA in many organisms.
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* Present address: Lexicon Genetics Incorporated, 4000 Research Forest Drive, The Woodlands, TX 77381
To whom reprint requests should be addressed at: Building 539/Room 243, National Cancer Institute, Frederick, MD 21702. E-mail: court@ncifcrf.gov. <<
Emphasis mine. Science fictional, no? They are discussing, in PNAS, DNA repair in humans. Wowsers!
doubletwist.com
I keep thinking this sort of stuff Kling profiles should be getting commercialized. So far, I've only noticed one, APBI's new TempliPhi amplification system.
Cheers, Tuck |
