The present invention relates to screening methods for identification of agents (e.g., small molecules) that modulate cytotoxic T lymphocyte antigen-specific target (e.g., tumor) cell killing, as well as to uses of compounds identified thereby as immunomodulatory, including use of EGFR inhibitors as immunomodulatory agents.

A polynucleotide encoding a modified luciferase polypeptide. The modified luciferase polypeptide has at least 60% amino acid sequence identity to a wild-type Oplophorus luciferase and includes at least one amino acid substitution at a position corresponding to an amino acid in a wild-type Oplophorus luciferase of SEQ ID NO:1. The modified luciferase polypeptide has at least one of enhanced luminescence, enhanced signal stability, and enhanced protein stability relative to the wild-type Oplophorus luciferase.

The present invention provides, among other things, methods of quantitating mRNA capping efficiency, particularly for mRNA synthesized in vitro. In some embodiments, the methods comprise chromatographic methods of quantifying capping efficiency and methylation status of the caps.

Provided herein are substantially non-luminescent peptide/polypeptide tags that are inserted internally within a protein of interest or between N-terminal and C-terminal peptides/polypeptides. Interaction of the internally-inserted tag with a complement polypeptide/peptide that is also substantially non-luminescent results in the formation a bioluminescent reporter complex.

Provided are methods for detecting protein interactions in a sample, the methods comprising: (a) detecting two or more polypeptides that when associated emit a first detectable signal in a first light emission spectrum; (b) contacting the two or more polypeptides with a third polypeptide conjugated to a dipole acceptor moiety that has a second light emission spectrum when excited within a light excitation spectrum, wherein the light excitation spectrum overlaps with the first light emission spectrum; and (c) detecting a second detectable signal emitted in the second light emission spectrum by the dipole acceptor moiety. Also provided are bioluminescent complexes comprising: (a) a first polypeptide conjugated to a dipole acceptor moiety, wherein the emits a first detectable signal in a first light emission spectrum.

The present invention provides a polypeptide comprising an N-terminal polypeptide domain having luciferase activity and a C-terminal polypeptide domain having VAMP1, VAMP2 or VAMP3 activity where VAMP stands for vesicle-associated membrane protein. Corresponding nucleic acid molecules, expression vectors and genetically modified cells are also provided. The invention also provides methods and uses of the same.

The present invention relates to compositions comprising and methods of producing genetically engineered bacteriophages, bacteriophage-like particles and non-replicating transduction particles (NRTPs) that contain non-native tail fibers that display altered host specificity and/or reactivity. The present invention also relates to methods of using these bacteriophages and NRTPs for the development of novel diagnostics, therapeutics and/or research reagents for bacteria-related diseases.

The present invention embraces an RNA replicon (self-amplifying RNA vector (saRNA)) that can be replicated by a replicase of a self-replicating virus, e.g., a replicase of alphavirus origin. According to the invention, translation of the replicase open reading frame is uncoupled from a 5′-terminal cap by placing translation of the replicase open reading frame under the translational control of an internal ribosome entry site (IRES). Thereby the initiation of translation depends on the molecular properties of the respective IRES, which compared to cap-dependent translation may require less or no cellular initiation factors to direct the ribosome to the translational start site. According to the invention, IRES-controlled replicase translation may allow the use of uncapped synthetic saRNA. Furthermore, the use of an IRES provides for the option to insert additional transgenes upstream to the IRES.

This disclosure describes recombinant angiotensin-converting enzyme II (ACE2) polypeptides, fusion proteins, and compositions thereof having improved binding affinity for the SARS-CoV-2 spike protein receptor binding domain relative to wild-type ACE2. Also provided are methods of using the recombinant ACE2 polypeptides, fusion proteins, and compositions thereof for treating subjects infected with a SARS-CoV-2 virus (i.e., subjects with COVID-19), subjects having symptoms suggestive of a SARS-CoV-2 infection, and subjects exposed to or at risk of exposure to SARS-CoV-2 virus. Other virus infections may also be treated.

Improved formulations of lipid nanoparticles are provided. Use of the lipid nanoparticles for delivery of a therapeutic agent and methods for their preparation are also provided.