Epicentre Technologies Corporation

Cellscript, LLC

University of Pennsylvania

Illumina Corporation

Other

University of Texas System

Promega Corporation

Jerome J Jendrisak

Gary A Dahl

Anthony Person

Judith Meis

Haiying Li Grunenwald

Nicholas Caruccio

Drew Weissman

Katalin Kariko

Ramesh Vaidyanathan

Rui Sousa

Ronald Meis

Leslie M Hoffman

Cynthia Chin

Robert E Smith

Kevin L Gunderson

Molly Min He

Frank J Steemers

Sasan Amini

Amirali Kia

Christian Gloeckner

Trina Faye Osothprarop

Agnes Radek

Michael J Fiandt

Fiona Kaper

Martin Ken Lewis

Igor Yu Goryshin

Mark A Maffitt

Natasha Pignatelli

Thomas Royce

Darin J Haskins

Elena K Davydova

Lucia B Rothman-Denes

Katalin Karikó

Svetalana Y Gerdes

Anthony D Person

Casey Turk

Reza Kahlor

Joanne Decker

Cynthia S Chin

Kia Amirali

Reza Kalhor

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

Presented herein are methods and compositions for tagmentation of nucleic acids. The methods are useful for generating tagged DNA fragments that are qualitatively and quantitatively representative of the target nucleic acids in the sample from which they are generated.

Transposase compositions for reduction of insertion bias

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').

Method of sequential tagmentation with transposase compositions for reduction of insertion bias

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

Jerome J Jendrisak