Monsanto Technology LLC

Ervin D Nagy

Xudong Ye

Jaishree M Chittoor

Stanislaw Flasinski

Elysia K Krieger

Krishnakumar Sridharan

Linda Rymarquis

Richard J Lawrence

Andrei Y Kouranov

Brent Brower-Toland

Veena Veena

Rosemarie Kuehn

Keith H Turner

Larry A Gilbertson

John D Bradley

Christine Shyu

Peizhen Yang

David G Caldwell

The present disclosure provides methods to accelerate recombination at selected genomic loci, allowing recombination to occur, and selecting events with molecular variation within the selected loci. The accelerated recombination generates novel variations in gene clusters that are present in the plant or mammalian genomes.

Enhanced recombination of genomic loci

Provided herein are systems, methods, and compositions for the modification of target DNA sequences. More particularly, systems, methods, and compositions for cleaving a target DNA in eukaryotic cells with a guide RNA capable of hybridizing with a target sequence and an RNA-guided DNA nuclease are provided. Also provided are vectors and vector systems which encode one or more components of a CRISPR complex, as well as methods for the design and use of such vectors. Also provided are methods for identifying and validating novel CRISPR systems.

RNA-guided DNA nucleases and uses thereof

This disclosure provides compositions and methods for improving site-directed integration of nucleic acids using RNA-guided nucleases coupled with HUH endonucleases. In one aspect, this disclosure provides a ribonucleoprotein comprising: (a) a re-combinant polypeptid comprising: (i) an amino acid sequence encoding a Cas12a nuclease; (ii) an amino acid sequence encoding a linker; and (iii) an amino acid sequence encoding a HUH nuclease; and (b) at least one guide nucleic acid.

Methods and compositions to promote targeted genome modifications using HUH endonucleases

This disclosure is related to plant-optimized recombinant nucleic acids encoding Cpf1 and their use in planta. Also disclosed are compositions, expression cassettes, and plant cells comprising the recombinant nucleic acids as well as methods and kits for modifying a target sequence in a plant genome using the recombinant nucleic acids.

Compositions and methods for genome editing in planta

The present disclosure relates to compositions and methods for improving site-directed integration frequency in the genome of plant cells.

Methods to improve site-directed integration frequency

Provided herein are systems, methods, and compositions for the modification of target DNA sequences. More particularly, systems, methods, and compositions for editing genomic DNA in eukaryotic cells with a CRISPR-associated transposase are provided. Also provided are vectors and vector systems which encode one or more CRISPR-associated transposases, as well as methods for the design and use of such vectors. Also provided are methods for identifying and validating novel CRISPR-associated transposases.

CRISPR-associated transposases and uses thereof

The disclosure provides novel corn, tomato, and soybean U6, U3, U2, U5, and 7SL snRNA promoters which are useful for CRISPR/Cas-mediated targeted gene modifications in plants. The disclosure also provides methods for use for U6, U3, U2, U5, and 7SL promoters in driving expression of sgRNA polynucleotides which function in a CRISPR/Cas system of targeted gene modification in plants. The disclosure also provides methods of genome modification by insertion of blunt-end DNA fragments at a site of genomic cleavage.

Compositions and methods for site directed genomic modification

This disclosure provides an engineered system comprising: a first nucleic acid molecule encoding a CasX nuclease, and a guide RNA (gRNA) or a second nucleic acid molecule encoding the gRNA, where the first nucleic acid molecule is codon optimized for a eukaryotic cell, and where the gRNA is designed to hybridize with a target site in the eukaryotic cell. Further, this disclosure provides a method of modifying at least one target site in a eukaryotic genome comprising: providing a eukaryotic cell with a CasX nuclease or a first nucleic acid molecule encoding the CasX nuclease, and providing the eukaryotic cell with a guide RNA (gRNA) or a second nucleic acid molecule encoding the gRNA, where the gRNA and the CasX nuclease form a complex, where the gRNA hybridizes to the target site, and where the complex generates a modification at the target site.

Growing community of inventors

St. Louis, MO, United States of America

Ervin D Nagy