USE
The present invention relates to the use of one or more cas genes for modulating resistance in a cell against a target nucleic acid or a transcription product thereof.
1 .- 86 . (canceled)
132 . A construct comprising:
a) a nucleic acid sequence encoding at least one clustered-regularly-spaced-short-palindromic-repeat-associated (cas) protein;
b) a nucleic acid sequence comprising at least one Clustered Regularly Spaced Short Palindromic Repeat (CRISPR) repeat or a full length complement thereof; and,
c) a nucleic acid sequence comprising at least one CRISPR spacer or a full length complement thereof.
133 . The construct of claim 132 , wherein expression of said construct in a cell results in modifying a nucleic acid of interest in the genomic DNA of said cell.
134 . The construct of claim 132 , comprising at least two CRISPR repeats.
135 . The construct of claim 132 , wherein the at least one cas protein and at least one CRISPR repeat are derived from the same organism.
136 . The construct of claim 132 , wherein the at least one cas protein and at least one CRISPR repeat are derived from the same cell.
137 . The construct of claim 132 , wherein the at least one CRISPR spacer is a synthetic nucleic acid.
138 . The construct of claim 132 , wherein the at least one CRISPR spacer is derived from an organism that is different to the organism from which the at least one cas protein and at least one CRISPR repeat are derived.
139 . The construct of claim 132 , wherein the at least one CRISPR spacer has at least 70% nucleotide sequence identity to a nucleic acid of interest in the genomic DNA of a cell.
140 . The construct of claim 132 , wherein the cas protein is encoded by a nucleic acid sequence having 90% nucleotide sequence identity to a nucleic acid sequence of SEQ ID NO: 462, 474, 489, 494, 499, 505 or 518.
141 . A vector comprising a construct of claim 132 .
142 . A host cell comprising a construct of claim 132 .
143 . A recombinant CRISPR-cas combination comprising at least one cas protein encoded by a cas gene and at least one CRISPR repeat in combination with at least one CRISPR spacer.
144 . The recombinant CRISPR-cas combination of claim 143 , wherein at least one cas gene and the combination of the at least one CRISPR repeat and at least one CRISPR spacer are on separate nucleic acids.
145 . The recombinant CRISPR-cas combination of claim 143 , wherein the combination comprises at least two CRISPR repeats.
146 . The recombinant CRISPR-cas combination of claim 143 , wherein the at least one cas gene or protein and at least one CRISPR repeat are derived from the same organism.
147 . The recombinant CRISPR-cas combination of claim 143 , wherein the at least one cas gene or protein and at least one CRISPR repeat are derived from the same cell.
148 . The recombinant CRISPR-cas combination of claim 143 , wherein the at least one CRISPR spacer is a synthetic nucleic acid.
149 . The recombinant CRISPR-cas combination of claim 143 , wherein the at least one CRISPR spacer is derived from an organism that is different to the cell from which the at least one cas gene and at least one CRISPR repeat are derived.
150 . The recombinant CRISPR-cas combination of claim 143 , wherein the CRISPR spacer is heterologous to the at least one cas gene and heterologous to the at least one CRISPR repeat.
151 . The recombinant CRISPR-cas combination of claim 143 , wherein the at least one CRISPR spacer has at least 70% nucleotide sequence identity to a nucleic acid of interest in the genomic DNA of a cell.
152 . The recombinant CRISPR-cas combinations of claim 143 , wherein the nucleotide sequence encoding the cas protein is at least 90% identical to a nucleic acid sequence of SEQ ID NO: 462, 474, 489, 494, 499, 505 or 518.
153 . A host cell comprising any one of the recombinant CRISPR-cas combinations of claim 143 .
154 . A method of modifying a nucleic acid of interest in the genomic DNA of a cell, the method comprising introducing into said cell at least one cas gene or protein and at least one CRISPR repeat in combination with at least one CRISPR spacer.
155 . The method of claim 154 wherein the at least one cas gene or protein and at least one CRISPR repeat are derived from the same organism.
156 . The method of claim 154 wherein the at least one cas gene or protein and at least one CRISPR repeat are derived from the same cell.
157 . The method of claim 154 wherein the at least one CRISPR spacer is a synthetic nucleic acid.
158 . The method of claim 154 wherein the at least one CRISPR spacer is derived from an organism that is different to the cell from which the at least one cas genes or proteins and at least one CRISPR repeat are derived.
159 . The method of claim 154 , wherein the at least one CRISPR spacer has at least 70% nucleotide sequence identity to a nucleic acid of interest in the genomic DNA of a cell.
160 . The method of claim 154 , comprising at least two CRISPR repeats.
161 . The method of claim 154 , wherein at least one cas gene and the combination of the at least one CRISPR repeat and at least one CRISPR spacer are on separate nucleic acids.
162 . The method of claim 154 further comprising evaluating the genomic DNA of said cell for a mutation.
163 . A recombinant nucleic acid comprising at least 50% of the wild type nucleotide sequence of SEQ ID NO: 462, 474, 489, 494, 499, 505 or 518.
164 . The recombinant construct of claim 163 comprising at least 65%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% of the wild type nucleotide sequence of SEQ ID NO: 462, 474, 489, 494, 499, 505 or 518.
165 . A recombinant nucleic acid comprising a nucleotide sequence of at least 75% sequence identity to SEQ ID NO: 462, 474, 489, 494, 499, 505 or 518.
166 . The recombinant construct of claim 165 comprising a nucleotide sequence of at least 85%, 90%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 462, 474, 489, 494, 499, 505 or 518.