Cell-associated secretion-enhancing fusion proteins are disclosed that comprise a target protein binding domain and a transmembrane retention domain. Co-expression in a host cell of a fusion protein and a target protein of interest that is temporarily bound by the fusion protein leads to an increased level of target protein secreted from the host cell. The fusion proteins are engineered to be retained with the producing host cell, thus eliminating a non-natural component from the extracellular media of the host cell and simplifying purification of the target protein. Nucleic acid molecules encoding such fusion proteins are also disclosed for use in expressing the fusion proteins in host cells, for use in restoring lost or diminished cell functions, and for use in treating diseases characterized by a lost or diminished cell function. Methods and compositions comprising fusion proteins of the invention are disclosed for use in enhancing the level of co-expressed target proteins secreted from host cells.

Cell-associated secretion-enhancing fusion proteins are disclosed that comprise a target protein binding domain and a transmembrane retention domain. Co-expression in a host cell of a fusion protein and a target protein of interest that is temporarily bound by the fusion protein leads to an increased level of target protein secreted from the host cell. The fusion proteins are engineered to be retained with the producing host cell, thus eliminating a non-natural component from the extracellular media of the host cell and simplifying purification of the target protein. Nucleic acid molecules encoding such fusion proteins are also disclosed for use in expressing the fusion proteins in host cells, for use in restoring lost or diminished cell functions, and for use in treating diseases characterized by a lost or diminished cell function. Methods and compositions comprising fusion proteins of the invention are disclosed for use in enhancing the level of co-expressed target proteins secreted from host cells.

A range of therapeutic mRNA molecules expressible to provide a target polypeptide or protein. The RNA molecules can contain one or more 5-methoxyuridines and 5-methylcytidines. Further provided are DNA templates, which can be transcribed to provide a target mRNA, and can have altered nucleotides, such as reduced deoxyadenosines. Also provided are processes for making the therapeutic mRNA molecules. The RNA molecules can be translated in vitro or in vivo to provide an active polypeptide or protein. The RNA molecules can be included in a composition used for preventing, treating, or ameliorating at least one symptom of a disease or condition in a subject in need thereof.

A range of therapeutic mRNA molecules expressible to provide a target polypeptide or protein. The RNA molecules can contain one or more 5-methoxyuridines and 5-methylcytidines. Further provided are DNA templates, which can be transcribed to provide a target mRNA, and can have altered nucleotides, such as reduced deoxyadenosines. Also provided are processes for making the therapeutic mRNA molecules. The RNA molecules can be translated in vitro or in vivo to provide an active polypeptide or protein. The RNA molecules can be included in a composition used for preventing, treating, or ameliorating at least one symptom of a disease or condition in a subject in need thereof.

This disclosure provides methods and compositions for treating TTP based on transfusion of a relatively small number of genetically modified red blood cells. The genetically modified red blood cells express a fusion protein including a fragment of ADAMTS13 that is enzymatically active against von Willebrand factor (VWF). The fragments of ADAMTS13 can be resistant to the inhibitors, e.g., the auto-immune antibodies, which are responsible for the acquired form of TTP.

This disclosure provides methods and compositions for treating TTP based on transfusion of a relatively small number of genetically modified red blood cells. The genetically modified red blood cells express a fusion protein including a fragment of ADAMTS13 that is enzymatically active against von Willebrand factor (VWF). The fragments of ADAMTS13 can be resistant to the inhibitors, e.g., the auto-immune antibodies, which are responsible for the acquired form of TTP.

The present invention relates to a transgenic cloned pig for xenotransplantation in which porcine endogenous retrovirus (RUN) EnvC is negative, α1,3-galactosyltransferase (GGTA1), CMP-N-acetylneuraminic acid hydroxylase (CMAH), isoglobotrihexosylceramide synthase (iGb3s), and beta-I,4-N-acetyl-galactosaminyl transferase2 (β4GalNT2) are knocked out, and human CD46 and thrombomodulin (TBM) genes are expressed, and to a method of preparing the transgenic cloned pig. The transgenic cloned pig according to the present invention may overcome hyperacute and antigen-antibody mediated immune rejection reaction, immune rejection reaction due to blood coagulation, and immune rejection reaction due to complement activity, without causing transfer of porcine endogenous retrovirus that occurs in xenotransplantation. Therefore, the transgenic cloned pig according to the present invention may be usefully utilized as a donor animal for xenotransplantation of organs and cells.

The present application discloses methods for synthetic production and cell-free synthesis of single stranded adeno-associated virus (AAV) vectors, for delivery and expression of a transgene in host cells. The present invention also relates to an in vitro process for production of closed-ended DNA vectors and corresponding single stranded AAV DNA vector products synthesized from the closed-ended DNA vectors having nicks.

The present application discloses methods for synthetic production and cell-free synthesis of single stranded adeno-associated virus (AAV) vectors, for delivery and expression of a transgene in host cells. The present invention also relates to an in vitro process for production of closed-ended DNA vectors and corresponding single stranded AAV DNA vector products synthesized from the closed-ended DNA vectors having nicks.

The two devices described allow for greater control over application of a fibrin biopolymer and yet remain simple enough to be used by both medical personnel and by non-medical workers. One of the devices includes chambers where components necessary for formation of a fibrin biopolymer are stored and are pushed by plungers connected to an actuator upon the actuator receiving pressure from the user, with the components mixing in a chamber within the device and flowing out of a tube connected to the mixing chamber before formation of the fibrin biopolymer is completed. The other device dispenses formulations that include components necessary for formations of a fibrin biopolymer onto a skin of a patient using one or more propellants, with the formation of the fibrin biopolymer being initiated after the formulations are dispensed on the patient's skin and are intentionally mixed together.