A lentiviral vector system for expressing a lentiviral particle is disclosed. The lentiviral vector system includes a therapeutic vector. The therapeutic vector comprises a phenylalanine hydroxylase (PAH) sequence for expressing at least one of PAH or a variant thereof, wherein the PAH sequence is truncated.

A method of constructing a hepatic precursor-like cell line is provided. The deposited number of the hepatic precursor-like cell line is CCTCC NO: C2019120. The construction method includes using primary hepatocytes as seed cells to establish the hepatic precursor-like cell line, which combines with controlling the conditions of passage culture, subculture, virus infection, proliferation culture and confluence culture. Moreover, a hepatocyte-specific transcription factor, FOXA3, is expressed in the immortalized cell line to make the hepatic precursor-like cell line differentiate easily. An application of the hepatic precursor-like cell line in the field of the bioartificial liver is also provided.

Identification of effective targets alleviating the programmed cell removal (PrCR) of tumor cells by macrophages is of very high interest. The present inventors have identified that the cyclin-dependent kinase inhibitor p21 protein is a strong regulator of the macrophage-mediated PrCR. Also, they showed that the adoptive transfer of p21 overexpressing monocytes induces macrophage PrCR and transition from an anti-inflammatory to a pro-inflammatory phenotype in vivo, delays cancer progression and increases significantly the overall survival of mice engrafted with cancer cells. The present invention therefore concerns therapeutic compositions comprising monocytes that over-express the cyclin-dependent kinase inhibitor p21 protein, and their use for treating mammals suffering from cancer, especially leukemia.

The present invention provides a chimeric antigen receptor, which includes: an extracellular domain, a transmembrane domain, and an intracellular domain connected in sequence. The extracellular domain includes an antigen recognition region; the intracellular domain includes a costimulatory signaling region and a CD3ζ intracellular region that are connected in sequence, to form a costimulatory signaling region-CD3ζ intracellular region; and the costimulatory signaling region-CD3ζ intracellular region is a polypeptide formed by mutation of lysine in a wild-type costimulatory signaling region-CD3ζ intracellular region into arginine. The present invention provides a method for optimization and modification of CAR-T, in which all lysine sites in an intracellular segment of CAR are mutated into arginine, thereby blocking ubiquitination modification of the CAR after antigen challenge. This strategy is applicable to different CARs and changing different intracellular costimulatory domains, and in particular provides a solution to the problem of poor proliferation of CAR-T in solid tumors.

The present invention relates to a method for preparation of an immortalized stem cell line that has an immortalizing gene introduced thereinto and retains an expression potential of the introduced gene while being restrained from proliferation so that the immortalized stem cell line can be used as a cell therapeutic agent, and to a use thereof. The immortalized stem cell line of the present invention, which has undergone a radiation process, cannot proliferate while maintaining the expression of the introduced foreign protein at a certain level or higher and as such, can be advantageously used as clinical samples such as various cell therapeutic agents.

Provided is an engineered immune cell comprising on its surface a recognition molecule that comprises a binding moiety specifically binding to a target molecule on the surface of a target cell, wherein the target molecule comprises an extracellular domain, and wherein the immune cell is capable of killing a target cell that comprises on its surface the target molecule. In one aspect, the binding moiety specifically binds to a distal portion of the extracellular domain, and the immune cell is capable of killing a target cell that comprises on its surface both the target molecule and the recognition molecule. In another aspect, the binding moiety specifically binds to a proximal portion of the extracellular domain, and the engineered immune cell has no or reduced capability of killing a target cell comprising on its surface both the target molecule and the recognition molecule.

Provided are anti-CD4 immune cell receptors that comprise an extracellular domain comprising a CD4 binding moiety that specifically binds to an epitope within a certain domain of CD4, a transmembrane domain, and an intracellular signaling domain. Also provided are engineered immune cells comprising such anti-CD4 immune cell receptors and uses thereof.

The present invention provides compositions and methods for transducing cells (e.g. T cells or immune cells). Also provided herein are methods of treating a disease in a subject in need thereof.

A process for producing blood coagulation Factor VII in large scale in 3 human cell lines (HepG2, Sk-Hep, and HKB-11) and to select the best recombinant protein producer is described. The murine line BHK-21 was used as control. The data allowed for the assertion that the system used to modify cell lines was efficient, so that all the cells were satisfactorily modified, and produced the protein of interest in a stable form. In addition, when comparing the murine line BHK-21 with the human cells (HepG2, Sk-Hep-1 and HKB-11), the latter proved to be able to produce rFVII more efficiently, which allows us to conclude that human cell lines are a great alternative to produce recombinant blood coagulation factors in large scale.

The present invention relates to compositions and methods for activating expression from the paternally-inherited allele of UBE3A in human Angelman's Syndrome neurons using viral vector delivery of short hairpin RNAs, ribozymes, and/or microRNAs.