The Scripps Research Institute

Leidos, Inc.

Stephen A Chappell

Vincent P Mauro

Gerald M Edelman

Geoffrey Owens

Wei Zhou

Frederick S Jones

Robyn Meech

John Dresios

Steven Gerald Haupt

Leslie Krushel

Jason K Pinkstaff

An array-based system of assembled DNA for computer data storage is described. An array surface contains immobilized seed DNA initially having with blunt (or blocked) ends with a photocleavable optical linker at a forward end thereof holding the last few base pairs. A light source is light is applied to break the linker, generating a sticky end which allows for hybridization. Data-bearing DNA cassettes are introduced to the array and attach via their sticky ends to the unblock sites on the array surface. The attachment is made permanent via ligase.

Optical methods and systems for DNA assembly for computer data storage

The present disclosure provides materials and methods for cell-free expression of epitopes for immunotherapy applications. In particular, the present disclosure provides materials and methods for expressing concatenated epitopes using a cell-free protein synthesis platform for high throughput, large scale, and unbiased epitope screening and the generation of multi-epitope vaccines.

Materials and methods for cell-free expression of vaccine epitope concatemers

Described herein are rules to modify natural mRNAs or to engineer synthetic mRNAs to increase their translation efficiencies. These rules describe modifications to mRNA coding and 3' UTR sequences intended to enhance protein synthesis by: 1) decreasing ribosomal diversion via AUG or non-canonical initiation codons in coding sequences, and/or 2) by evading miRNA-mediated down-regulation by eliminating one or more miRNA binding sites in coding sequences.

Reengineering mRNA primary structure for enhanced protein production

Synthetic and isolated translational regulatory elements, including oligonucleotides that have translational enhancing activity, internal ribosome entry site (IRES) activity, or translational inhibitory activity, and multimers of such translational regulatory elements are provided. In addition, compositions that include such translational regulatory elements are provided, as are methods of using the translational regulatory elements.

Synthetic internal ribosome entry sites and methods of identifying same

Provided are methods of identifying oligonucleotides having transcriptional or translational activity by integrating ilie oligonucleotide into a eukaryotic cell genome such that the oligonucleotide is operatively linked to an expressible polynucleotide, and detecting a change in expression of the expressible polynucleotide due to the operatively linked oligonucleotide. Also provided are vectors useful for identifying an oligonucleotide having transcriptional or translational regulatory activity according to a method of the invention. In addition, isolated synthetic transcriptional or translational regulatory elements identified according to a method of the invention are provided, as are kits, which contain a vector useful for identifying a transcriptional or translational regulatory element, or an isolated synthetic transcriptional or translational regulatory element or plurality of such elements. Also provided are isolated transcriptional regulatory elements.

Methods of identifying synthetic transcriptional and translational regulatory elements, and compositions relating to same

Provided are methods of identifying oligonucleotides having transcriptional or translational activity by integrating the oligonucleotide into a eukaryotic cell genome such that the oligonucleotide is operatively linked to an expressible polynucleotide, and detecting a change in expression of the expressible polynucleotide due to the operatively linked oligonucleotide. Also provided are vectors useful for identifying an oligonucleotide having transcriptional or translational regulatory activity according to a method of the invention. In addition, isolated synthetic transcriptional or translational regulatory elements identified according to a method of the invention are provided, as are kits, which contain a vector useful for identifying a transcriptional or translational regulatory element, or an isolated synthetic transcriptional or translational regulatory element or plurality of such elements. Also provided are isolated transcriptional regulatory elements.

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Del Mar, CA, United States of America

Stephen A Chappell