Filamentous fungal mutants with improved homologous recombination efficiency
AssigneeDsm IP Assets B.V.
InventorsPetrus Jacobus Theodorus Dekker, Marco Alexander van den Berg
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Abstract
The present invention relates to a method for increasing the efficiency of targeted integration of a polynucleotide to a pre-determined site into the genome of a filamentous fungal cell with a preference for NHR, wherein said polynucleotide has a region of homology with said pre-determined site, comprising steering an integration pathway towards HR. The present invention also relates to a mutant filamentous fungus originating from a parent cell, said mutant having an HR pathway with elevated efficiency and/or an NHR pathway with a lowered efficiency and/or a NHR/HR ratio with decreased efficiency as compared to said HR and/or NHR efficiency and/or NHR/HR ratio of said parent cell under the same conditions.
Claims14
A mutant of a parent filamentous fungal cell, the parent organism having a preference for non-homologous recombination (NHR), wherein the filamentous fungal host cell is a Penicillium species, wherein the ratio of NHR/homologous recombination (HR) is decreased in the mutant as compared to said ratio in said parent organism measured under the same conditions, wherein the mutant is deficient in a gene encoding a component involved in NHR, and/or has a decreased level of a component involved in NHR, wherein the mutant is, optionally inducibly, deficient in at least one filamentous fungal homologue of yeast KU80, yeast KU70, or both, or has, optionally inducibly, a decreased amount of at least one of the proteins encoded by these genes.
A method for increasing the efficiency of targeted integration of a polynucleotide to a pre-determined site into the genome of a filamentous fungal cell with a preference for NHR, wherein said polynucleotide has a region of homology with said pre-determined site, wherein said filamentous fungal cell belongs the genus Penicillium, comprising providing a mutant of a parent filamentous fungal cell, wherein the ratio of NHR/HR is decreased in the mutant as compared to said ratio in said parent organism measured under the same conditions, wherein said mutant is deficient in a gene encoding a component involved in NHR, and/or has a decreased level of a component involved in NHR, wherein the mutant is, optionally inducibly, deficient in at least one filamentous fungal homologue of yeast KU80, yeast KU70, or both, or has, optionally inducibly, a decreased amount of at least one of the proteins encoded by these genes.
Dependencies are inferred from the claim text.