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    Preparation of Combinatorial Libraries of DNA Constructs
    1.
    发明申请
    Preparation of Combinatorial Libraries of DNA Constructs 有权

    公开(公告)号：US20210301285A1

    公开(公告)日：2021-09-30

    申请号：US17265092

    申请日：2019-07-19

    Applicant: Novozymes A/S

    Inventor： Michael Lynge Nielsen ,  Nicholas Jochumsen ,  José Arnau

    IPC: C12N15/10 ,  C12N15/81 ,  C12N15/90

    Abstract: Means and methods for preparing combinatorial libraries of DNA constructs, in particular expression cassettes, including nucleic acid constructs, expression vectors, host cells, methods for preparing host cells, and methods for producing polypeptides of interest, whereby the expression comprises an first intron and a second intron on either side of the polynucleotide to be expressed and a promoter and a terminator. Also claimed is a method of constructing eukaryotic host cells in which the cells are contacted with three polynucleotides and in which the first and second and the second and third are pairwise capable of homologous recombination and of subsequent formation of introns.

    Modified Filamentous Fungal Host Cells
    2.
    发明申请
    Modified Filamentous Fungal Host Cells 有权

    公开(公告)号：US20220025423A1

    公开(公告)日：2022-01-27

    申请号：US17296938

    申请日：2019-11-26

    Applicant: Novozymes A/S

    Inventor： Nicholas Jochumsen ,  Michael Rey ,  Chiho Inoue

    IPC: C12P21/02 ,  C12N9/04 ,  C12N15/80

    Abstract: The present invention relates to mutated filamentous fungal host cell producing a secreted polypeptide of interest, wherein a native putative steroid dehydrogenase is modified, truncated, partly or fully inactivated, present at reduced level or eliminated compared to a non-mutated parent cell, and wherein said native putative steroid dehydrogenase comprises at least one conserved amino acid motif selected from: YGAR and/or VPHS[W/Y]F and/or QC[A/V/S]RRL and/or LKKYTLP and/or CPHYT, and methods of producing a secreted polypeptide of interest in said cells as well as methods of producing said cells.

    Genome Editing by Guided Endonuclease and Single-stranded Oligonucleotide
    4.
    发明申请
    Genome Editing by Guided Endonuclease and Single-stranded Oligonucleotide 有权

    公开(公告)号：US20220010305A1

    公开(公告)日：2022-01-13

    申请号：US17289953

    申请日：2019-10-30

    Applicant: Novozymes A/S

    Inventor： Nicholas Jochumsen ,  Gloria Muzzi-Erichsen

    IPC: C12N15/11 ,  C12N15/10

    Abstract: The present invention relates to methods for introducing one or more desired nucleotide modification(s) in a target sequence in the genome of a microorganism cell using a polynucleotide-guided endonuclease, e.g., the MAD7 enzyme isolated and described by Inscripta™ or the well-known Streptococcus pyogenes Cas9, together with a suitable guide RNA for each target sequence to be modified, to generate a site-specific cut or nick in at least one genome target sequence, followed by the repair of the cut(s) and/or nick(s) via at least one oligonucleotide capable of hybridizing with the at least one genome target sequence, thereby highly efficiently introducing the one or more desired modification(s) into the target sequence.

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