The present invention relates to chemically inducible split protein actuators (CISPA), which utilize ligand-binding proteins or protein domains originating from humans or other organisms, which are rationally split into two fragments that reassemble only in the presence of a cognate ligand. In particular, the invention relates to their design, manufacture, structure, and uses. The designed CISPAs can be used to regulate cellular processes such as gene expression, conditionally reconstitute of the function of a protein such as enzyme activity, as biological sensors, or for other applications.

This vector set is induced into host cells, and cells for producing heteromultimeric proteins are selected from the host cells into which has the vector set is introduced. vector set is a set of a first expression vector and a second expression vector, wherein an expression cassette of at least one subunit X which is a part of a subunit constituting a heteromultimeric protein is contained in both the first expression vector and the second expression vector and from among the subunits constituting the heteromultimeric protein, an expression cassette of the remaining subunit Y excluding the subunit X is contained for each type in one among the first expression vector and the second expression vector.

A method of characterizing a protein is provided herein. The method includes accessing simulated protein structure data with a computer system, where the simulated protein structure data indicate a structure of the protein. The method further includes quantifying, using the computer system and based on the simulated protein structure data, a plurality of dynamics metrics for a plurality of residues in the protein. The plurality of dynamics metrics are related to functional behaviors of the protein using the computer system. Additionally, the method includes generating a report from the functional behaviors and the dynamics metrics using the computer system, where the report comprises a functional characterization of each residue in the plurality of residues and a functional characterization of the protein.

Fusion proteins and methods for conditionally fine-tuning transposase protein stability and activity for clinical applications are provided herein

Various embodiments are disclosed herein relate to methods for selection of a genetically engineered cell. Some embodiments relate to a cell that is produced with the methods disclosed herein.

The invention provides a platform and methods of using the platform for the regulation of the expression of a target gene using exposure to an aptamer ligand (for example, a small molecule). The platform features a polynucleotide gene regulation cassette that is placed in the target gene and includes a synthetic riboswitch positioned in the context of a 5 intron-alternative exon-3 intron. The riboswitch comprises an effector region and a sensor region (e.g., an aptamer that binds a small molecule ligand) such that the alternative exon is spliced into the target gene mRNA when the ligand is not present thereby preventing expression of the target gene. When the ligand is present, the alternative exon is not spliced into the target gene mRNA thereby providing expression of the target gene.

The present disclosure relates to providing an engineered CHO cell line wherein the two essential metabolic genes are knocked out. Particularly, the present invention relates to a double knockout CHO cell line (DHFR/ and GS/) with disrupted Dihydrofolate Reductase (DHFR) and Glutamine Synthetase (GS) genomic loci. The double knockout CHO cell line (DHFR/ and GS/) being suitable for expression of monoclonal antibodies, dimeric therapeutic proteins, Fab, single chain fragments, or the like. The present disclosure also provides method of generation of a double knock out CHO cell line (DHFR/ and GS/) using gene selection and/or manipulating techniques such as CRISPR/Cas9 system, Zinc Finger Nuclease, TALEN, or the like. The present disclosure further provides method of selection of clones and production of therapeutic proteins of interest with increased titre.

The present disclosure relates to mutant polypeptides derived from Escherichia coli dihydrofolate reductase (DHFR) which can be fused to a polypeptide of interest for efficient conditional modulation of its activity. Also disclosed are polynucleotides encoding such mutant polypeptides, vectors comprising such polynucleotides, and the use of such polypeptides, polynucleotides and vectors for treating a disorder.

The present disclosure reports that a calcium-dependent serine protease, complement component is (C1s) has been identified as a protease responsive for cleavage of exogenous polypeptides expressed in mammalian cell lines such as CHO cells. These CHO cell lines provide for increased yield of antigenically correct, uncleaved exogenous polypeptide as compared to unmodified CHO cells expressing the active C1s protease. C1s-deficient cell lines and methods for use of same for producing exogenous polypeptides, e.g., human immunodeficiency virus (HIV) envelope glycoprotein polypeptides, such as, gp120 or human Factor VIII are provided.

Disclosed are composition comprising (a) an inducible transgene construct, comprising a sequence encoding an inducible promoter and a sequence encoding a transgene, and (b) a receptor construct, comprising a sequence encoding a constitutive promoter and a sequence encoding an exogenous receptor, wherein, upon integration of the construct of (a) and the construct of (b) into a genomic sequence of a cell, the exogenous reporter is expressed, and wherein the exogenous reporter, upon binding a ligand, transduces an intracellular signal that targets the inducible promoter of (a) to modify gene expression. Methods for introducing compositions into cells and the use of the resultant cells in adoptive cell therapies are also provided.