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January 23, 2014
Clicked-DNA Works In Human Cells
I made reference to the (surprising) ability of artificially-linked "click" DNA analogs (an earlier example here) to show activity in cells when Ali Tavassoli (of U. Southampton) spoke about his lab's work back at the Challenges in Chemical Biology conference last summer.
Here's their 2012 paper on the subject, and now they have another one out in Angewandte Chemie, which extends the work to human cells. It sort of boggles me mind to think that these things are actually transcriptionally active, but they're lighting up human cells with the fluorescent dye mCherry, and doing (from what I can see) all the appropriate control experiments. (It's not being reworked by repair enzymes along the way, for example). Here's the wrap-up:
The practical limit on the length of error-free oligonu- cleotide synthesis has necessitated the use of enzymes for the assembly of polynucleotide chains into genes. However, these approaches have been constrained by the assumption that the phosphodiester backbone that links oligonucleotides is critical for the biocompatibility and cellular function of the resulting DNA. As demonstrated in this work, this is not the case. Our results strongly suggest that RNA polymerase II, the enzyme responsible for all mRNA synthesis in eukaryotes, correctly transcribes the genetic information contained on a click-linked strand of DNA. . .Our results indicate that a phosphodiester linker is not essential for joining oligonucleotides for gene synthesis and open up the possibility of replacing enzymatic ligation with highly efficient chemical reactions. This approach would not necessarily be limited to the linker reported here, and alternative chemical reactions and the resulting linkers may also be suitable for this purpose.
I look forward to seeing what use chemical biology will make of this sort of thing. Now, can you make functional mRNAs out of this as well?
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