Compositions and methods disclosed herein relate to improved in vitro cell-free systems for various biological reactions. In one aspect, a composition of the present disclosure includes: a cell-free extract derived from an organism; a nucleic acid; and an organelle that is heterologous to the organism.

The present invention relates to: a method for producing immunocytes, specifically induced natural killer T (iNKT) cells that are induced by direct reprogramming of isolated somatic cells, and chimeric antigen receptor (CAR)-iNKT cells into which a CAR gene encoding a CAR is introduced; iNKT cells produced by the method; and a cell therapy composition and a pharmaceutical composition for preventing or treating cancer, comprising the iNKT cells.

The method according to the present invention can produce, through direct reprogramming, iNKT cells or iNKT cells into which a CAR gene is introduced, from isolated cells so as to simplify the production process and shorten production time, thereby reducing costs, to have excellent NKT cell production efficiency, and to ensure safety according to the production without passing through induced pluripotent stem cells, thereby having an excellent NKT cell production effect distinguished from that of a conventional reprogramming technique. In addition, the iNKT cells or iNKT cells into which a CAR gene is introduced, which are produced by the method, have an excellent cancer cell killing ability, and thus can be effectively used as a cell therapy composition or a pharmaceutical composition for preventing or treating cancer.

The present application relates to a chimeric antigen receptor (CAR) which comprises a target-dependent on-switch CAR. The CAR of the invention may reduce cytotoxicity towards normal cells and improve CAR-T safety. CAR molecules were designed using the transmembrane and juxtamembrane motifs of the IL2 receptor β chain (IL2Rβ or IL2Rb), the L ow-Density Lipoprotein Receptor (LDLR), the Seizure 6-like Protein 2 (SEZ6L2), and degradation sequence (PSKFFSQL) of IL2Rβ, which resulted in greatly reduced CAR expression at the cell surface in the absence of target antigen, while retaining downstream activation ability in response to antigen-expressing target cells. In the absence of target antigen, CAR surface expression is undetectable. The present application has shown that primary T cells expressing these surface-unstable CAR variants are able to elicit antigen-dependent target cell killing. By limiting CAR activity in this way, the present application can reduce therapeutic toxicity and T cell exhaustion. Due to its limited detectability in the absence of antigen, the present application refers to this system as a “Stealth CAR”. The present application further relates to compositions, preparation methods and uses of the Stealth CAR of the present application.

Real-time systems for monitoring base editing in living cells, including base editing by APOBEC-Cas9 fusions, is provided herein.

New insecticidal proteins, nucleotides, peptides, their expression in plants, methods of producing the peptides, new processes, production techniques, new peptides, new formulations, and new organisms, a process which increases the insecticidal peptide production yield from yeast expression systems. The present invention is also related and discloses selected endotoxins we call cysteine rich insecticidal peptides (CRIPS) which are peptides derived from Bacillus thuringiensis (Bt) and their genes and endotoxins in combination with toxic peptides known as Inhibitor Cystine Knot (ICK) genes and peptides as well as with other types of insecticidal peptides such as trypsin modulating oostatic factor (TMOF) peptide sequences used in various formulations and combinations; of both genes and peptides, useful for the control of insects.

The invention relates to novel means and processes for the detection of a virus in a biological sample comprising cells infected by the virus. In particular, the invention relates to a fluorescent reporter protein designed as a recombinant inactive form of flipGFP suitable for specific activation by viral components in particular by viral proteins, such as viral protease, wherein the viral component recognizes a cleavage site inserted in the recombinant flipGFP. The fluorescent reporter protein is suitable for use in an in vitro method of detection of virus infection in a biological sample when the virus is related to the viral components activating the inactive form of flipGFP into an active fluorescent flipGFP in a biological sample, especially a sample comprising cells, in particular unaltered cells.

Compositions for characterization of botulinum toxin (BoNT) are described that include a genetically modified cell that is transfected with an artificial construct comprising a nucleic acid sequence that encodes for a hybrid protein having (a) a reporter-containing portion chemically coupled to (b) a cleavage site and (c) a control fluorophore. The cleavage site interacts with a BoNT in a manner that cleaves the reporter-containing portion from remainder of the construct. The cleaved portion is destroyed or otherwise degraded by the local environment, and presence of BoNT is evidenced by reduction in signal from the reporter. The cleavage sequence is all or part of a SNARE protein, the cleavable reporter-containing portion is preferably Yellow Fluorescent Protein (YFP), Citrine, Venus, or a YPet protein and the control fluorophore is preferably CFP, mStrawberry, or a mCherry protein.

Disclosed are immunogenic proteins from Pseudomonas aeruginosa as well as nucleic acids, vectors and transformed cells useful for expression of the proteins. Also disclosed are methods for prophylaxis of infection with Pseudomonas aeruginosa using the proteins, nucleic acids, vectors or transformed cells.

A polypeptide that comprises the amino acid sequence of SEQ ID NO: 1 and has a function of delivering a substance into cells, a polypeptide that comprises the amino acid sequence of SEQ ID NO: 2 and has a function of degrading a nucleic acid molecule, a polypeptide that comprises the amino acid sequence of SEQ ID NO: 3 and has a function of degrading a nucleic acid molecule and a function of intranuclearly migrating the same, polynucleotides encoding the respective polypeptides, and transformed cells containing the respective polynucleotides.

Described herein is a composition for delivery of an active agent. The composition includes a peptide coacervate, wherein the peptide coacervate includes one or more peptides derived from histidine-rich proteins, and an active agent encapsulated in the peptide coacervate. Further provided are a method for encapsulation of an active agent in a peptide coacervate, a method for delivery of an active agent, and a method for treating or diagnosing a condition or disease in a subject in need thereof.