GSJ: Received Jul. 25, 2007: http://wbabin.net/saba/saba88.htm

Display technologies are extremely valuable in protein science, and involve associating an encoding nucleic acid with its protein product (1).

Herein are novel means of mRNA display, involving noncovalent association of a portion of a mRNA, with a portion of its protein product.

A preferred embodiment is shown in Figure 1.

The individual mRNAs need be sufficiently separated to favor association of a mRNA with its product. To favor this end, notice the common aptamer-binding portions of the proteins are synthesized prior to the different portions, such that the aptamers have the potential of capturing the proteins before their complete synthesis. Since such a capture would be intramolecular, its rate would be expected to be far greater than intermolecular capture. In certain situations it may be desirable to perform translation in micro, nano, or pico wells; or micro, nano or pico vessels.

Some important variations of Figure 1 include, positioning the aptamers within or at the 5’ ends the mRNAs, positioning the aptamer-binding portions of the proteins other than at the N-termini, and replacing the aptamer-binding portions of the proteins with unnatural amino acids (2).

The common aptamers in Figure 1 could be replaced by an RNA structure (3,4) or modification (5) which functions as a ligand for common portions of the proteins. Figure 2 exemplifies this derivation using an unnatural nucleotide.

The great advantage of this display technology over phage display is that translation can be done in vitro, and thus unnatural nucleotides and unnatural amino acids can be easily incorporated, greatly expanding the diversity of the libraries synthesized.

If it should be that the above invention is indeed novel any patentable rights I may have, I freely give away.

It is hoped that others will honor the invention as delineated above and by the following claims.

Claims

References (incorporated in their entirety by reference)

1) Display technologies: application for the discovery of drug and gene delivery agents.
Sergeeva, et al Adv Drug Deliv Rev. 2006 Dec 30;58(15):1622-54. Epub 2006 Oct 1

2) Expanding the genetic code.
Wang, et al Annu Rev Biophys Biomol Struct. 2006;35:225-49

3) RNA-protein complexes.
Cusack S Curr Opin Struct Biol. 1999 Feb;9(1):66-73

4) RNA-binding proteins: modular design for efficient function.
Lunde, et al Nat Rev Mol Cell Biol. 2007 Jun;8(6):479-90

5) Site-specific biotinylation of RNA molecules by transcription using unnatural base pairs.
Moriyama, et al Nucleic Acids Res. 2005 Aug 19;33(15):e129

6) Cell-free protein expression and functional assay in nanowell chip format.
Angenendt, et al Anal Chem. 2004 Apr 1;76(7):1844-9

7) Toxin detection by a miniaturized in vitro protein expression array.
Mei, et al Anal Chem. 2005 Sep 1;77(17):5494-500

8) Single-molecule PCR using water-in-oil emulsion.
Nakano, et al J Biotechnol. 2003 Apr 24;102(2):117-24

9) PCR amplification from single DNA molecules on magnetic beads in emulsion: application for high-throughput screening of transcription factor targets.
Kojima, et al Nucleic Acids Res. 2005 Oct 6;33(17):e150

Addendum Jul 28, 2007

Like phage display, and unlike covalent in vitro display systems, this noncovalent technology allows for multiple successive screening against a target. That is after each screening those mRNA selected are simply retranslated prior to reexposure to the target.

Addendum Jul 30, 2007

A variation of the process in Figure 2 would be to use mRNAs wherein each had multiple interaction ligands as shown in Figure 3.