The present invention relates to a process for constructing a single vector comprising cas9 and sgRNA (all-in-one vector) for mycobacterial genome modifications, comprising the steps of (a) providing the expression cassette of cas9 and the tracrRNA expression cassette with codons optimized for expression in mycobacteria and clone into a cloning vector; (b) sequentially cloning the sequences optimized in step (a) into a mycobacterial bridge vector; (c) selecting targeting sequences (crRNAs) specific to the gene of interest; (d) inserting the crRNAs selected in step (c) into the mycobacterial expression vector, upstream of the tracrRNA sequence, at position +1 downstream of an inducible promoter for the construction of the sgRNA cassette; and (e) adding at the end of each expression cassette a transcriptional terminator.

A CRISPR-based genome editing is disclosed. More specifically, nucleic acids, compositions and kits for editing a human HBD gene and their use in the treatment of haemoglobinopathies are disclosed. Methods of making thereof and methods of editing a human HBD gene are also disclosed. The composition and/or kit contains (a) a first component selected from a guide RNA (gRNA) or a DNA polynucleotide encoding the gRNA, (b) a second component selected from a DNA donor template or a vector containing the DNA donor template, and (c) a third component containing a Cas9 polypeptide or a variant thereof or a nucleic acid encoding a Cas9 polypeptide or a variant thereof.

An anti-CD371 (anti-CLL-1) chimeric antigen receptor (CAR), engineered immune cells comprising the CAR, as well as therapeutic compositions, therapeutic methods and companion diagnostic methods are disclosed herein.

The invention relates to methods of forward programming an induced pluripotent stem cell (iPSC) into a somatic cell, said methods comprising at least dual targeting of safe harbour sites in the genome of an iPSC. The invention also includes cells obtained by such methods.

The present disclosure includes agents that specifically bind CLL-1, such as antibodies and chimeric antigen receptors, as well as methods of making and using such agents.

Described herein is a method of fungal population modification via inter-domain conjugation, including co-culturing a bacterial population and the fungal population under growth conditions for the bacteria, and maintaining growth of the bacterial population during the co-culturing by controlling an essential nutrient for growth of the bacterial population. A bacterial plasmid is transferred to at least a portion of the fungal population to provide the inter-domain modification.

Recombinant DNA molecules and constructs are provided that are useful for modulating gene expression in plants. One or more expression cassette(s) of a recombinant DNA molecule or construct may be excised from transgenic plants following transformation by the presence of flanking site-specific recombination sites in the recombinant DNA molecule or construct by expression of a site-specific recombinase enzyme encoded by the recombinant DNA molecule or construct. Such a recombinase system may be used to remove expression cassette(s) from plants transformed with the recombinant DNA construct or vector. The recombinase transgene may be operably linked to a promoter suitable for autoexcision in transformed plants without crossing to a different transgenic line expressing the recombinase. Methods for causing autoexcision of one or more expression cassette(s) in a transgenic plant, and plants and cells containing or transformed with a recombinant DNA molecule or construct of the present disclosure, are also provided.

The invention relates to a bacterial cell comprising a Cas1 RT fusion protein, a Cas2 protein and a CRISPR direct repeat (DR) sequence, wherein an RNA polymerase promoter in addition to the leader sequence is associated with the DR sequence. The invention further relates to a composition comprising two bacterial cell populations, each comprising a Cas1 RT fusion protein and Cas2 protein. The two cell types contain different versions of a CRISPR direct repeat (DR) sequence. The invention further relates to methods for analysis of transcription recording events of bacteria having passed through a subject's intestine, to assign a probability to the subject having a condition, such as malnutrition or inflammation of the intestine.

Compositions and methods for inserting a nucleic acid encoding a product of interest into a genomic safe harbor locus in a cell, a population of cells, or a subject or for expressing a nucleic acid encoding a product of interest from a genomic safe harbor locus in a cell, a population of cells, or a subject are provided. Also provided are cells or populations of cells comprising a nucleic acid construct comprising a coding sequence for a product of interest inserted into a genomic safe harbor locus. Also provided are methods of identifying genomic safe harbor loci for use in specific cell or tissue types.

Provided herein are systems of engineered Class 2, Type V nucleases and guide ribonucleic acid scaffolds useful for the editing of target nucleic acids. Also provided are methods of making and using such systems to modify nucleic acids.