Methods and compositions for targeted gene modification
AssigneesCibus US LLC, Cibus Europe B.V.
InventorsPeter R. Beetham, Gregory F. W. Gocal, Christian Schopke, Noel Sauer, James Pearce, Rosa E. Segami, Jerry Mozoruk
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Abstract
Provided herein include methods and compositions for effecting a targeted genetic change in DNA in a cell. Certain aspects and embodiments relate to improving the efficiency of the targeting of modifications to specific locations in genomic or other nucleotide sequences. As described herein, nucleic acids which direct specific changes to the genome may be combined with various approaches to enhance the availability of components of the natural repair systems present in the cells being targeted for modification.
Claims17
A method of causing a targeted genetic change within a plant cell, comprising:
- contacting the plant cell with (i) a gene repair oligonucleobase (GRON) configured to mediate introduction of the targeted genetic change within the gene, and (ii) a Clustered Regularly Interspaced Short Palindromic Repeat complex configured to introduce single strand or double strand breaks within the gene (a CRISPR), under conditions in which the targeted genetic change is introduced within the plant cell,
- wherein the GRON configured to mediate introduction of the targeted genetic change within the gene comprises one or more alterations from conventional nucleic acids selected from the group consisting of one or more 2′O-methyl nucleotides at the 5′ end thereof, one or more 2′O-methyl nucleotides at the 3′ end thereof, a terminus cap at the 5′ end thereof, a fluorescent dye conjugated at the 5′ end thereof, a fluorescent dye conjugated to the 3′ end thereof, one or more phosphothioate residues at the 5′ end thereof, one or more phosphothioate residues at the 3′ end thereof, and a reverse base at the 3′ end thereof, and, wherein the targeted genetic change is introduced without the insertion of the GRON configured to mediate introduction of the targeted change within the gene.
Dependencies are inferred from the claim text.