The present disclosure pertains to compositions comprising aflibercept and methods for producing such compositions in chemically defined media and using chromatography to reduce amounts of certain aflibercept variants.

The present invention relates to endoglucanase variants with improved properties and uses thereof as eco-friendly biocatalysts in various industrial processes. More in particular the invention provides a polypeptide with endoglucanase activity (EC 3.2.1.4) comprising an amino acid sequence that is at least 90% identical to the amino acid sequence according to SEQ ID NO: 1 or SEQ ID NO: 2, wherein the polypeptide comprises an amino acid selected from the group consisting of Glycine, Threonine, Serine, Alanine, Leucine, Isoleucine and Valine at a position corresponding to position 579 in SEQ ID NO: 1 or SEQ ID NO: 2 or an amino acid selected from the group consisting of Serine, Alanine and Valine at a position corresponding to position 656 in SEQ ID NO: 1 or SEQ ID NO: 2.

The present disclosure discloses a visualized screening method for an Aspergillus recombinant strain with multigene editing and belongs to the technical field of gene engineering. CRISPR-Cas9 is used in the disclosure to cleave spore color change-related genes and a target gene in Aspergillus at the same time, such that editing of the target gene is visualized and an Aspergillus niger strain with multigene editing can be rapidly and efficiently screened out through spore phenotypes. Through different combinations of visualized genes and non-phenotypic change genes, rapid screening of the strain with multigene editing and simultaneous screening of multiple visualized genes are realized, and use of resistance genes in industrial strains is reduced.

The instant invention relates to a filamentous fungal host cell producing a polypeptide of interest, said host cell comprising one or more native or heterologous polynucleotide encoding a long non-coding RNA (lncRNA), wherein said lncRNA comprises more than 262 nucleotides and comprises a contiguous nucleotide sequence at its 3′ end, absent any poly-adenylation tail, that is at least 70% identical to that of SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO: 10, SEQ ID NO: 19, SEQ ID NO:20 or SEQ ID NO:21; and/or said lncRNA comprises 2 or more Xyr1 binding sequences, as well as methods of producing a polypeptide of interest in said host cells and methods of improving the production of a polypeptide of interest.

The invention provides hybrid and recombinant phagemid vectors for expressing a transgene in a target cell transduced with the vector. A recombinant phagemid particle comprises at least one transgene expression cassette which encodes an agent which exerts a biological effect on the target cell, characterised in that the phagemid particle comprises a genome which lacks at least 50% of its bacteriophage genome. The invention extends to the use of such phagemid expression systems as a research tool, and for the delivery of transgenes in a variety of gene therapy applications, DNA and/or peptide vaccine delivery and imaging techniques. The invention extends to in vitro, in vivo or in situ methods for producing viral vectors, such as recombinant adeno-associated viruses (rAAV) or lentivirus vectors (rLV), and to genetic constructs used in such methods.

The present disclosure pertains to compositions comprising anti-VEGF proteins and methods for producing such compositions.

The disclosure discloses an Aspergillus niger engineered strain for reducing byproduct fumaric acid in a fermentation process of L-malic acid. The Aspergillus niger engineered strain is an Aspergillus niger engineered strain in which a fumarate hydratase gene fum is knocked out. The disclosure overcomes the defects in the prior art, in the current process of producing malic acid through fermentation of Aspergillus niger, byproduct fumaric acid can be accumulated with the generation of malic acid so as to cause the improved cost of the subsequent malic acid purification process. The disclosure provides an Aspergillus niger engineered strain in which a fum gene is knocked out and a method for greatly reducing byproduct fumaric acid in the fermentation production of Aspergillus niger.

This invention relates to recombinant nucleic constructs comprising CRISPR-Cas effector proteins, reverse transcriptases and extended guide nucleic acids and methods of use thereof for modifying nucleic acids in plants.

Described herein are synthetic inducible promoters including 5′-(UP element)-(−35 element)-(spacer element)-(−10 element)-(discriminator element)-3′. Also included are vectors, α-Proteobacteria strains and γ-Proteobacteria strains including the synthetic inducible promoters. A Mobile-CRISPRi plasmid and methods of partially or fully knocking-down expression of a gene in α-Proteobacteria or γ-Proteobacteria are also described. Further included are methods of making an α-Proteobacteria or γ-Proteobacteria strain.

The disclosure relates to modification of a heat resistant cytoplasmic heat stable 6-phosphogluconate dehydrogenase (6PGDH) enzyme by fusing the cytoplasmic 6PGDH enzyme in frame to a plastid-targeting sequence. This modification allows the import of the cytoplasmic 6PGDH enzyme into plastids of a plant cell. Polynucleotides encoding and expressing the modified cytoplasmic 6PGDH enzymes are provided. The disclosure further provides transgenic plants and seeds containing the disclosed polynucleotides and expressing the modified cytoplasmic 6PGDH enzymes during development. The invention further relates to methods for developing a transgenic plant that has increased heat resistance and yield during heat stress.