Disclosed are methods of determining activity of CDK4 and CDK6 variants upon exposure to CDK inhibitors, methods for determining activity of a Rb variant, methods for determining the activity of a p16 variant in a cell, and methods for determining the sensitivity of a CDK4 variant or a CDK6 variant to p16 in a cell. Stable cell lines for determining activity of CDK4 variants, CDK6 variants, Rb variants, and p16 variants are also disclosed.

Circular handed alpha-helical repeat proteins are described. The repeat proteins have a number of uses as scaffolds for geometrically precise, arrayed presentation of cell-signaling or immune-related protein and peptide epitopes, as well as numerous other therapeutic, diagnostic, and nanotechnological uses.

Described herein are methods and compositions for genomic editing. Endonucleases for genomic editing involve inducing breaks in double stranded DNA, for which knock-ins are notoriously inefficient for relying on random integration of homologous DNA sequences into the break site by repair proteins. To address these issues, described herein are novel recombinant fusion proteins that actively recruit linear DNA inserts in closer proximity to the genomic cleavage site, increasing integration efficiency of large DNA fragments into the genome. Such improvements to genomic editing technology allow one to use lower linear DNA concentrations without sacrificing efficiency and can be further combined with other features, such as fluorescent protein reporting systems.

The present invention addresses the problem of providing: a fusion protein or conjugated protein having excellent cell membrane permeability, containing a partial peptide derived from human, and suitable for intracellular delivery; an intracellular delivery carrier comprising such a fusion protein or conjugated protein; a partial peptide; a cell membrane permeation enhancer comprising the partial peptide; DNA; and a vector. The fusion protein or conjugated protein has a partial peptide comprising at least seven consecutive amino acid residues of an amino acid sequence encoded by a predetermined DNA, and a ligand directly or indirectly bound to the partial peptide and having the capability of binding to cell surfaces. The ligand is preferably an antibody. The intracellular delivery carrier comprises the fusion protein or conjugated protein. The cell membrane permeation enhancer comprises the partial peptide.

The present disclosure provides a genetically encoded calcium indicator (GECI), nucleic acids encoding the GECI, and host cells comprising the GECI. The present disclosure also provides methods of detecting a change in the intracellular concentration of a cell expressing a GECI of the present disclosure.

The present invention provides a method for controlling colocalization of two or more proteins in a cell. The method comprises expressing the proteins, scaffold RNA molecules having binding motifs for the proteins, and a trigger RNA molecule in the cell. In the presence of the trigger RNA molecule, a scaffold may be assembled (ON) by the scaffold RNA molecules via hybridization such that the proteins may be colocalized; or disassembled (OFF) such that the proteins may be separated and not colocalized. The proteins may provide a biological activity when colocalized or not colocalized.

The present invention is directed to compositions comprising modified SMARCB1 and uses thereof.

The present invention provides a recombinant cell for producing para-nitro-L-phenylalanine (pN-Phe). The recombinant cell comprises heterologous genes encoding heterologous enzymes. The recombinant cell expresses the heterologous enzymes and contains a native metabolite. The native metabolite is converted to the pN-Phe in the recombinant cell. The biosynthesized pN-Phe may be incorporated into a target polypeptide in the recombinant cell without requiring exposure of the recombinant cell to exogenous pN-Phe. A cell culture comprising the recombinant cell is also provided. Further provided is a method of producing pN-Phe by a recombinant cell comprising heterologous genes encoding heterologous enzymes. The method comprises expressing a native metabolite by the recombinant cell, expressing the heterologous enzymes, and converting the native metabolite to the pN-Phe in the recombinant cell. The method may further comprise incorporating the pN-Phe into the target polypeptide in the recombinant cell.

The present invention provides a method for detecting and assaying Clostridium neurotoxins and identification of serotypes of botulinum neurotoxins in various food matrices and clinical samples. This method is also used for detection of BoNT inside the neuronal and epithelial cells. The method comprises detecting and assaying the presence of a Clostridium neurotoxin in a sample by: exposing the sample containing a Clostridium neurotoxin to a sample comprising a novel SNAMPXIN/SNAMP universal recombinant substrate fusion protein capable of producing a detectable FRET, following cleavage; detecting and assaying the presence of the Clostridium neurotoxin by measuring a change in the energy transfer or the luminescence signal; and detecting and assaying an electrophoretic mobility pattern of one or more cleaved protein bands or a degraded protein, using a high throughput automated system to identify the different serotypes of the Clostridium neurotoxin. SNAMPXIN/SNAMP is formed from parts of BoNT substrates SNAP-25 and VAMP.

The present invention provides DNA constructs for producing transgenic plants. The DNA constructs encode a transit peptide that is targeted to the mitochondria and the chloroplasts within a cell. Also provided are plant cells and vectors comprising the DNA constructs. Methods of generating a transgenic plant using the vectors and constructs provided herein and methods of targeting a protein to mitochondria or chloroplasts in a cell.