Agilent Technologies, Inc.

University of Colorado

Leland Stanford Junior University

Lievre Cornu, LLC

Hewlett-packard Company

Lieure Cornu LLC

University and Agilent Technologies, Inc.

Douglas J Dellinger

Marvin H Caruthers

Joel Myerson

Geraldine F Dellinger

Agnieszka Sierzchala

Zoltan Timar

Daniel E Ryan

Robert Kaiser

Jason R Betley

Geraldine Fulcrand

Zoltan Kupihar

Mark Andrew Troll

Jeffrey Robert Sampson

Steven M Lefkowitz

Michel G M Perbost

Matthew Hebden Porteus

Rasmus O Bak

SueAnn C Dahm

Ayal Hendel

Victor R Mokler

Mithun Roy

Joe Clark

Luca Monfregola

Brian Phillip Smart

Charles Z Hotz

Diane D Ilsley

Paul A Metz

Benjamin D Lunstad

Fernando Ferreira

John Turner

Gary B Gordon

Robert A Ach

Bill J Peck

John S Hargreaves

Subhadeep Roy

Michael G M Perbost

Kenneth W Hill

Michel G M Perobost

Jeffrey R Sampson

Thomas Jacob Cobb

Eric LeProust

Richard Bacon

Logan Garner

John W Davies

Brian Stell

Matthew Williamson

Geraldino Dellinger

David James Neep

Agnieska Sierzchala

Peter Cogan

Agnieszka B Sierzchala

Mithum Roy

Geraldine F Dellingen

Paul Metz

Victor Molker

M Agnieszka Sierzchala

A resin formed of copolymer beads is disclosed. The copolymer beads have a first repeating motif, a second repeating motif having a substituted styrene with an ester functional group or an amide with a terminal amine or hydroxyl functional group, and a crosslinking agent having a divinyl compound. The copolymer beads are capable of expanding substantially uniformly in polar and non-polar solvents.

Polymer beads for oligonucleotide synthesis

Provided herein are compositions and methods for inducing CRISPR/Cas-based editing of a target nucleic acid (e.g., target DNA or target RNA) in vitro or in a cell, using modified prime editing guide RNAs (pegRNAs) that incorporate one or more chemically-modified nucleotides. The modified pegRNAs disclosed herein may be used to induce Cas-mediated incorporation of one or more nucleotide changes and/or targeted mutagenesis of a target nucleic acid. The nucleotide change can include, e.g., one or more nucleotide changes, an insertion of one or more nucleotides, or a deletion of one or more nucleotides.

Guide RNAs with chemical modification for prime editing

Provided herein are methods for inducing CRISPR/Cas-based gene regulation (e.g., genome editing or gene expression) of a target nucleic acid (e.g., target DNA or target RNA) in a cell. The methods include using modified single guide RNAs (sgRNAs) that enhance gene regulation of the target nucleic acid in a primary cell for use in ex vivo therapy or in a cell in a subject for use in in vivo therapy. Additionally, provided herein are methods for preventing or treating a genetic disease in a subject by administering a sufficient amount of a modified sgRNA to correct a mutation in a target gene associated with the genetic disease.

Compositions comprising chemically modified guide RNAs for CRISPR/Cas-mediated editing of HBB

Aspects of the present disclosure include compositions that make use of phosphorus and/or nucleobase protecting groups which find use in the synthesis of long polynucleotides. Phosphorus protecting groups are provided that help increase the stepwise coupling yield and/or phosphorous protecting groups that can be removed during the oxidation step. Amidine nucleobase protecting groups are provided that find use in the subject compositions and methods which provides for e.g., increased resistance to depurination during polynucleotide synthesis. In some instances, the methods and compositions disclosed herein utilize a combination of the phosphorus and amidine nucleobase protecting groups in the synthesis of polynucleotides having a sequence of 200 or more monomeric units in length. Also provided are methods for synthesizing a polynucleotide (e.g., a DNA) using one or more compounds disclosed herein.

Phosphorous protecting groups and methods of preparation and use thereof

Compounds and methods for purifying oligonucleotides such as RNA and DNA. A target oligonucleotide is reacted with an orthoester linker comprising an affinity tag to form an orthoester oligonucleotide-orthoester linker conjugate which is subjected to a purification technique to separate the target oligonucleotide from impurities such as truncated oligonucleotides. The orthoester linker can be then removed under mild conditions to generate the target oligonucleotide in high purity.

Orthoester compositions for affinity purification of oligonucleotides

Chemically modified guide RNAS for CRISPR/Cas-mediated gene regulation

Chemically modified guide RNAs for CRISPR/CAS-mediated gene regulation

The present invention relates to modified guide RNAs and their use in clustered, regularly interspaced, short palindromic repeats (CRISPR)/CRISPR-associated (Cas) systems.

Guide RNA with chemical modifications

The present invention relates to guide RNAs having chemical modifications and their use in CRISPR-Cas systems. The chemically modified guide RNAs have enhanced specificity for target polynucleotide sequences. The present invention also relates to methods of using chemically modified guide RNAs for cleaving or nicking polynucleotides, and for high specificity genome editing.

High specificity genome editing using chemically modified guide RNAs

A method of deprotecting a solid support bound polynucleotide comprising at least one 2'-protected ribonucleotide in which a step of contacting the polynucleotide with a composition comprising a diamine is performed under conditions sufficient to deprotect and cleave the polynucleotide which remains retained on the solid support.

Protected monomer and method of final deprotection for RNA synthesis

A method of deprotecting a solid support bound polynucleotide includes the step of contacting the polynucleotide with a composition comprising a diamine under conditions sufficient to deprotect the 2'-protected ribonucleotide residue. The solid support bound polynucleotide has at least one 2&#x2032;-protected ribonucleotide residue, which has the following structure: wherein Bis a protected or unprotected heterocycle; Ris a protecting group selected from a hydrocarbyl, a substituted hydrocarbyl, an aryl, and a substituted aryl; X is O or S; and PG is a thionocarbamate protecting group.

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Boulder, CO, United States of America

Douglas J Dellinger