Changes for the better

Eisenstein, Michael

doi:10.1038/nmeth1104-180

Classic Protocol
Published: November 2004

Changes for the better

Michael Eisenstein

Nature Methods volume 1, page 180 (2004)Cite this article

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“What sort of molecule could function as a specific mutagen?” pondered Joshua Lederberg in his 1958 Nobel lecture. “How could this be generally accomplished except by another molecule of conforming length and periodicity, that is an analogous polynucleotide?” This genetics luminary recognized the nature of the problem but the absence of techniques for gene sequencing and the generation of complimentary oligonucleotides remained a serious obstacle, and Lederberg concluded that “pending such advances, specific mutagenesis is an implausible expectation.” Fortunately, this would not remain the case for long.

In 1971, Clyde A. Hutchison III was a newly appointed faculty member at the University of North Carolina, studying the single-stranded DNA bacteriophage ΦX174. In one early paper, Hutchison and colleague Marshall Edgell demonstrated the 'salvage' of mutant phage by annealing mutant genomic DNA with enzymatically cleaved fragments from wild-type phage¹; these findings suggested the possibility of DNA sequence modification, but without sequence data and access to targeted oligonucleotides of appropriate length, this technique's potential could not be realized.

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