Recorded: 22 Mar 2003
So here I was studying bacteriophage and I knew a lot about phage. You know we worked with phage genetics and did biochemistry on it. It was accessible; you could manipulate it. We did wonderful things with the Lambda repressor in the DNA, but you simply couldn’t do that in higher cells because of the complexity in the genome. So the question is how could you do that and the answer to that was by recombinant DNA. But when we started nothing existed. There was no method. There was no way to do it. The only thing that had been done at the time was that somebody took a piece of frog DNA, spliced it into a plasmid and put into bacteria.
That was in 1971, I think and it was [Stanley] Cohen and [Herbert] Boyer in, of course, the famous Stanford patent came out of that.
And so we were really starting from scratch. And the first and most obvious thing to do is to clone cDNA. And it was known that you could extend mRNA with, using an oligo dT primer to make the first strand, but that was it. And so with Arg [Argiris Efstratiadis], who I’ve mentioned before we worked out conditions for making full-length copies and then used a tailing method that had actually been worked out at Stanford to insert the cDNAs.
And we did that with an eye towards getting full-length cDNAs. Other groups were working on it at the time and in fact one group published before us. But it was a tiny bit of DNA and it wasn’t particularly useful. And so when we finally put all the pieces together which was, you know, actually two papers preceded that, and we finally finished it, it was tremendous. It was really, really exciting because now we had established a method that one could use to isolate and clone a copy of any messenger RNA in a cell. And, of course, the importance of that is obvious now. I mean everything we do is through cDNA cloning.
It was in 1975, 1976, I think. We first published a Cell paper on making full-length cDNA transcript, then we had a paper on full-length synthesis of double stranded—
Mila Pollock: Just can you give me dates?
Well, the first, you know, the first cDNA paper was in April of ’75, the second paper was in February of ’76 was the full-length synthesis of double stranded DNA. And the actual cloning of cDNA was published in June of 1976.
Tom Maniatis, molecular biologist, is a leader in the field of recombinant DNA. At Vanderbilt University he completed his Ph.D. studying DNA wide-angle scattering. He became a postdoctoral fellow and professor at Harvard University and met Jim Watson just before he became director of Cold Spring Harbor Laboratory.
While Maniatis was beginning experimentation with cDNA cloning and gene regulation of higher cells, the controversy over recombinant DNA in Cambridge stunted his progression. Watson offered Maniatis a position at CSHL where he could work more efficiently to understand the methods of recombinant DNA. At CSHL, Maniatis completed full-length synthesis of double stranded DNA and actual cloning of cDNA.
He is currently a professor of molecular and cellular biology at Harvard University studying the mechanisms involved in the regulation of RNA transciption and pre-messenger RNA splicing. He studies transcription to understand how eukaryotic genes are activated by viral infection and extracellular signals.