Epicentre Technologies Corporation

Cellscript, LLC

University of Pennsylvania

Other

Illumina Corporation

Gary A Dahl

Jerome J Jendrisak

Anthony Person

Judith Meis

Haiying Li Grunenwald

Nicholas Caruccio

Drew Weissman

Katalin Kariko

Cynthia Chin

Ramesh Vaidyanathan

Ronald Meis

Roy Rabindranauth Sooknanan

Agnes Radek

Robert E Smith

Elena K Davydova

Lucia B Rothman-Denes

Michael J Fiandt

Katalin Karikó

Svetalana Y Gerdes

Anthony D Person

Joanne Decker

Cynthia S Chin

The present invention provides methods, compositions and kits for using a transposase and a transposon end for generating extensive fragmentation and 5'-tagging of double-stranded target DNA in vitro, then using a DNA polymerase for generating 5′- and 3′-tagged single-stranded DNA fragments without performing a PCR amplification reaction, wherein the first tag on the 5′-ends exhibits the sequence of the transferred transposon end and optionally, an additional arbitrary sequence, and the second tag on the 3′-ends exhibits a different sequence from the sequence exhibited by the first tag. The method is useful for generating 5′- and 3′-tagged DNA fragments for use in a variety of processes, including processes for metagenomic analysis of DNA in environmental samples, copy number variation (CNV) analysis of DNA, and comparative genomic sequencing (CGS), including massively parallel DNA sequencing (so-called 'next generation sequencing').

Transposon end compositions and methods for modifying nucleic acids

The present invention relates to compositions, kits and methods for making and using RNA compositions comprising in vitro-synthesized ssRNA inducing a biological or biochemical effect in a mammalian cell or organism into which the RNA composition is repeatedly or continuously introduced. In certain embodiments, the invention provides compositions and methods for changing the state of differentiation or phenotype of a human or other vertebrate cell. For example, the present invention provides mRNA and methods for reprogramming cells that exhibit a first differentiated state or phenotype to cells that exhibit a second differentiated state or phenotype, such as to reprogram human somatic cells to pluripotent stem cells.

Making and using in vitro-synthesized ssRNA for introducing into mammalian cells to induce a biological or biochemical effect

The present invention provides compositions and methods for reprogramming somatic cells using purified RNA preparations comprising single-strand mRNA encoding an iPS cell induction factor. The purified RNA preparations are preferably substantially free of RNA contaminant molecules that: i) would activate an immune response in the somatic cells, ii) would decrease expression of the single-stranded mRNA in the somatic cells, and/or iii) active RNA sensors in the somatic cells. In certain embodiments, the purified RNA preparations are substantially free of partial mRNAs, double-stranded RNAs, un-capped RNA molecules, and/or single-stranded run-on mRNAs.

RNA preparations comprising purified modified RNA for reprogramming cells

The present invention provides methods, compositions and kits for using a transposase and a transposon end for generating extensive fragmentation and 5'-tagging of double-stranded target DNA in vitro, then using a DNA polymerase for generating 5&#x2032;- and 3&#x2032;-tagged single-stranded DNA fragments without performing a PCR amplification reaction, wherein the first tag on the 5&#x2032;-ends exhibits the sequence of the transferred transposon end and optionally, an additional arbitrary sequence, and the second tag on the 3&#x2032;-ends exhibits a different sequence from the sequence exhibited by the first tag. The method is useful for generating 5&#x2032;- and 3&#x2032;-tagged DNA fragments for use in a variety of processes, including processes for metagenomic analysis of DNA in environmental samples, copy number variation (CNV) analysis of DNA, and comparative genomic sequencing (CGS), including massively parallel DNA sequencing so-called 'next generation sequencing').

The present invention provides methods, kits, and compositions for reducing an innate immune system response in a human or animal cell, tissue or organism. One embodiment comprises: introducing an Agent mRNA comprising in vitro-synthesized mRNA encoding one or more proteins that affect the induction, activity or response of an innate immune response pathway; whereby, the innate immune response in the cell, tissue or organism is reduced compared to the innate immune response in the absence of the Agent mRNA. Other embodiments are methods, compositions and kits for using an Agent mRNA for treating a disease or medical condition in a human or animal that exhibits symptoms of an elevated innate immune system, or for reducing an innate immune response that is induced in a human or animal cell, tissue or organism by a Foreign Substance that is administered to the cell, tissue or organism.

Inhibition of innate immune response

The present invention provides methods, compositions and kits for using a transposase and a transposon end for generating extensive fragmentation and 5'-tagging of double-stranded target DNA in vitro, then using a DNA polymerase for generating 5&#x2032;- and 3&#x2032;-tagged single-stranded DNA fragments without performing a PCR amplification reaction, wherein the first tag on the 5&#x2032;-ends exhibits the sequence of the transferred transposon end and optionally, an additional arbitrary sequence, and the second tag on the 3&#x2032;-ends exhibits a different sequence from the sequence exhibited by the first tag. The method is useful for generating 5&#x2032;- and 3&#x2032;-tagged DNA fragments for use in a variety of processes, including processes for metagenomic analysis of DNA in environmental samples, copy number variation (CNV) analysis of DNA, and comparative genomic sequencing (CGS), including massively parallel DNA sequencing (so-called 'next generation sequencing).

The present invention provides novel compositions, kits and methods employing RNA 5' polyphosphatases, RNA 5&#x2032; monophosphatases, capping enzymes, decapping enzymes, nucleic acid pyrophosphatases and RNA ligases, as well as other enzymes, for selective 5&#x2032; ligation tagging of desired classes of RNA molecules that differ with respect to particular chemical moieties on their 5&#x2032; ends. The 5&#x2032;  tagged RNA molecules can be used for synthesis of tagged first-stand cDNA, double-stranded cDNA, and sense or antisense RNA for a variety of uses.

Selective 5' ligation tagging of RNA

The present invention relates to methods for changing the state of differentiation of a eukaryotic cell, the methods comprising introducing mRNA encoding one or more reprogramming factors into a cell and maintaining the cell under conditions wherein the cell is viable and the mRNA that is introduced into the cell is expressed in sufficient amount and for sufficient time to generate a cell that exhibits a changed state of differentiation compared to the cell into which the mRNA was introduced, and compositions therefor. For example, the present invention provides mRNA molecules and methods for their use to reprogram human somatic cells into pluripotent stem cells.

Compositions and methods for reprogramming eukaryotic cells

Growing community of inventors

Madison, WI, United States of America

Gary A Dahl