RNA-guided programmable cytosine and adenine base editors are a powerful class of genome editing tool for the introduction of localized base transitions without generating a double-stranded DNA break. Base editors (BE) have an optimal window of activity relative to the PAM recognized by the Cas9 enzyme and these constructs are strand selective. Here we demonstrate that fusion of a programmable DNA-binding domain (pDBD) or another Cas9 orthologue to spCas9-BE, we can produce an RNA-programmable Cas9-BE-pDBD chimera or Cas9-BE-Cas9 chimeras with dramatically improved activities and increased targeting range. Cas9-pDBD or Cas9-Cas9 fusion base editors display an expanded targeting repertoire and achieve highly specific genome editing, which can be tailored to achieve extremely precise genome editing at nearly any genomic locus.

This invention pertains to optimized protein fusion linkers for creating multi-functional chimeric proteins and methods of using the same. Additionally, the invention pertains to chimeric proteins for use in guided endonuclease systems.

Sized-based detection techniques for detection of bio-nanoparticles are described. Detection nanoparticles and methods of forming the detection nanoparticles that can be utilized in the techniques are described. Detection nanoparticles can include modified bacteriophage that express a linking agent for a specific binding agent. Detection nanoparticles can bind a functional bio-nanoparticle with high specificity through specific binding of one or more entities unique to the functional bio-nanoparticle of interest. A detection nanoparticle can target an entity of a bio-nanoparticle that is relevant to its function, and as such, the methods can provide improvements in detection of complete and functional bio-nanoparticles. Size-based detection regimes can include particle displacement measurement techniques based upon Brownian motion.

Methods for manufacturing genetically engineered T cells expressing a chimeric antigen receptor (CAR), such as a CAR that binds human CD19, BCMA, or CD70, and having multiple additional gene edits, for example, a disrupted Regnase-1 gene, a disrupted TGFBRII gene, a disrupted TRAC gene, a disrupted β2M gene, or a combination thereof, using CRISPR/Cas gene editing systems.

An anti-CTLA4 (cytotoxic T lymphocyte associated antigen 4) and anti-PD-1 (programmed cell death 1) bifunctional antibody. a pharmaceutical composition thereof and use thereof. Particularly, the anti-CLTA4 and anti-PD-1 bifunctional antibody comprises a first protein functional domain that targets PD-1 and a second protein functional domain that targets CTLA-4. The bifunctional antibody can bind to CTLA-4 and PD-1 specifically, relieve immunosuppression of CTLA4 and PD-1 on an organism specifically, activate T lymphocytes, and thus has good application prospects.

An anti-CTLA4 (cytotoxic T lymphocyte associated antigen 4) and anti-PD-1 (programmed cell death 1) bifunctional antibody. a pharmaceutical composition thereof and use thereof. Particularly, the anti-CLTA4 and anti-PD-1 bifunctional antibody comprises a first protein functional domain that targets PD-1 and a second protein functional domain that targets CTLA-4. The bifunctional antibody can bind to CTLA-4 and PD-1 specifically, relieve immunosuppression of CTLA4 and PD-1 on an organism specifically, activate T lymphocytes, and thus has good application prospects.

The present invention relates to designed ankyrin repeat domains with altered surface residues, as well as to proteins comprising such a designed ankyrin repeat domain, nucleic acids encoding such domains or proteins, methods of preparing such proteins, pharmaceutical compositions comprising such proteins or nucleic acids, and the use of such proteins, nucleic acids or pharmaceutical compositions in the treatment of diseases.

A protein scaffold includes a plurality of EutM subunits and a multi-enzyme cascade. The multi-enzyme cascade includes a first enzyme attached to the first EutM subunit and a second enzyme attached to the second EutM subunit. The scaffold may be formed by a method that generally includes incubating a plurality of EutM subunits under conditions allowing the EutM subunits to self-assemble into a protein scaffold, attaching a first enzyme of a multi-enzyme cascade to a first EutM subunit, and attaching a second enzyme of the multi-enzyme cascade to a second EutM subunit. The scaffold may be self-assembled in vivo or in vitro. Each enzyme may be, independently of any other enzyme, attached to its EutM subunit in vivo or in vitro. Each enzyme may be, independently of any other enzyme, attached to its EutM subunit before or after the scaffold is assembled.

A protein scaffold includes a plurality of EutM subunits and a multi-enzyme cascade. The multi-enzyme cascade includes a first enzyme attached to the first EutM subunit and a second enzyme attached to the second EutM subunit. The scaffold may be formed by a method that generally includes incubating a plurality of EutM subunits under conditions allowing the EutM subunits to self-assemble into a protein scaffold, attaching a first enzyme of a multi-enzyme cascade to a first EutM subunit, and attaching a second enzyme of the multi-enzyme cascade to a second EutM subunit. The scaffold may be self-assembled in vivo or in vitro. Each enzyme may be, independently of any other enzyme, attached to its EutM subunit in vivo or in vitro. Each enzyme may be, independently of any other enzyme, attached to its EutM subunit before or after the scaffold is assembled.

The invention provides a novel adrenomedullin derivative sustainable for a long period which is capable of substantially suppressing unwanted side effects while maintaining pharmacological effects of adrenomedullin. An aspect of the invention relates to a compound represented by formula (I): [wherein A is an Fc region of an immunoglobulin, B is a peptide moiety derived from adrenomedullin or a modified form thereof with adrenomedullin activity, and L is a linking group comprising a peptide having any given amino acid sequence] or a salt thereof, or a hydrate thereof. Another aspect of the invention relates to a method for producing the compound represented by formula (I), and a medicament comprising the compound as an active ingredient.
A-L-B  (I).