Provided is a protein coded by a gene related to cell totipotency and a transcriptional activation domain of a mammalian YAP protein or a fusion protein of a segment with a transcriptional control activity, a coding nucleotide sequence, an expression vector and a composition thereof, as well as a method for inducing the pluripotent stem cells by using the fusion protein.

The present invention discloses a digital-to-analog conversion apparatus having signal calibration mechanism. A DAC circuit includes conversion circuits to generate an output analog signal and an echo-canceling analog signal. An echo transmission circuit performs signal processing on an echo path to generate an echo signal. An echo calibration circuit includes odd and even calibration circuits to perform mapping according to offset tables and perform processing according to response coefficients on odd and even input parts of an input digital signal to generate odd and even calibration parts of an echo-canceling calibration signal. A calibration parameter calculation circuit generates offsets according to an error signal between the echo signal and the echo-canceling calibration signal and path information related to the echo calibration circuit. The echo calibration circuit makes the coefficients converge according to the error signal and pseudo noise transmission path information, and updates the offset tables according to the offset.

An enveloped viral particle producer or packaging cell, wherein the cell is genetically engineered to decrease expression of MHC-I on the surface of the cell.

The present disclosure provides methods for genetically modifying lymphocytes and methods for performing adoptive cellular therapy that include transducing T cells and/or NK cells. The methods can include inhibitory RNA molecule(s) and/or engineered signaling polypeptides that can include a lymphoproliferative element, and/or a chimeric antigen receptor (CAR), for example a microenvironment restricted biologic CAR (MRB-CAR). Additional elements of such engineered signaling polypeptides are provided herein, such as those that drive proliferation and regulatory elements therefor, as well as replication incompetent recombinant retroviral particles and packaging cell lines and methods of making the same. Numerous elements and methods for regulating transduced and/or genetically modified T cells and/or NK cells are provided, such as, for example, those including riboswitches, MRB-CARs, recognition domains, and/or pH-modulating agents.

A cell for producing retroviral vectors comprising nucleic acid sequences encoding: i) gag-pol; ii) env; iii) the RNA genome of the retroviral vector; and iv) optionally rev, or a functional substitute thereof, wherein at least two nucleic acid sequences are located at the same genetic locus; and wherein the at least two nucleic acid sequences are in reverse and/or alternating orientations.

Human-derived virus-like particles (heVLPs), comprising a membrane comprising a phospholipid bilayer with one or more HERV-derived envelope proteins on the external side; one or more HERV-derived GAG proteins in the heVLP core, and a cargo molecule, e.g., a biomolecule and/or chemical cargo molecule, disposed in the core of the heVLP on the inside of the membrane, wherein the heVLP does not comprise a gag protein, except for gag proteins that are encoded in the human genome or gag proteins that are encoded by a consensus sequence that is derived from gag proteins found in the human genome, and methods of use thereof for delivery of the cargo molecule to cells.

Human-derived virus-like particles (heVLPs), comprising a membrane comprising a phospholipid bilayer with one or more HERV-derived envelope proteins on the external side; one or more HERV-derived GAG proteins in the heVLP core, and a cargo molecule, e.g., a biomolecule and/or chemical cargo molecule, disposed in the core of the heVLP on the inside of the membrane, wherein the heVLP does not comprise a gag protein, except for gag proteins that are encoded in the human genome or gag proteins that are encoded by a consensus sequence that is derived from gag proteins found in the human genome, and methods of use thereof for delivery of the cargo molecule to cells.

Embodiments provide systems and methods for transfecting cells to produce a viral vector or other selected virus. One embodiment of a method for transfecting cells to produce a selected virus includes providing a first solution comprising plasmids or other extrachromosomal DNA encoding the virus and a second solution comprising a transfection agent; the two solutions kept separate. The two solutions are then mixed to produce a transfection solution (TS) to be delivered to a cell culture medium (CCM) for transfection of cells in the medium with the plasmid DNA encoding the virus wherein the mixing is initiated responsive to a trigger event. The TS is then incubated to form transfection complexes. Subsequently, the TS is delivered to the CCM to transfect the cells in the medium wherein a viral production parameter resulting from transfection is optimized by initiating mixing of the solutions responsive to the trigger event.

An enveloped viral particle producer or packaging cell, wherein the cell is genetically engineered to decrease expression of CD47 on the surface of the cell.

The present disclosure provides methods for genetically modifying lymphocytes and methods for performing adoptive cellular therapy that include transducing T cells and/or NK cells. The methods can include inhibitory RNA molecule(s) and/or engineered signaling polypeptides that can include a lymphoproliferative element, and/or a chimeric antigen receptor (CAR), for example a microenvironment restricted biologic CAR (MRB-CAR). Additional elements of such engineered signaling polypeptides are provided herein, such as those that drive proliferation and regulatory elements therefor, as well as replication incompetent recombinant retroviral particles and packaging cell lines and methods of making the same. Numerous elements and methods for regulating transduced and/or genetically modified T cells and/or NK cells are provided, such as, for example, those including riboswitches, MRB-CARS, recognition domains, and/or pH-modulating agents.