Described herein are compositions, systems, and methods for obtaining primordial germ cells (PGCs) from pluripotent stem cells (PSCs). Inhibiting or bypassing tight junction formation in a population of pluripotent stem cells to generate a modified cell population, and contacting the modified cell population with BMP. Where inhibiting or bypassing tight junction formation includes incubating the population of pluripotent stem cells.

Compositions of previously uncharacterized thermostable class II, type VI CRISPR/Cas effector proteins are provided. The compositions include polynucleotides and vectors for expressing the effector proteins and one or more associated crRNAs, as well as cells that contain the effector proteins. Methods of preparation and use of the compositions are also disclosed. The compositions and methods are especially applicable to rapid and facile detection of nucleic acids in assays requiring elevated temperatures, RNA knockdown, RNA editing, antiviral activity, and RNA imaging.

Described herein are regulators of T cells as well as methods of modulating such T cell regulators, and methods of identifying new agents that modulate the T cell regulators. Modification of such T cell regulators in lymphoid and/or myeloid cells can provide lymphoid/myeloid cells that can be administered to subjects in need thereof, for example, subject suffering from immune disorders, cancer and other diseases and conditions.

Compositions and methods for ex vivo genomic editing using Neisseria meningitidis (Nine) CRISPR/Cas9 systems are disclosed. The present disclosure provides for engineered cells comprising a genetical modification for use e.g., in adoptive cell transfer therapies.

In some aspects, the present disclosure provides compositions comprising an inhibitory polynucleotide and either a guide polynucleotide and/or a polynucleotide that encodes for a nuclease or a nuclease; and a lipid nanoparticle comprising at least one ionizable lipid; wherein the each of the nucleic acids are encapsulated within the lipid nanoparticle, and pharmaceutical compositions thereof. The present disclosure also provides methods employing said compositions and/or pharmaceutical compositions, such as methods of treating diseases or disorders.

Methods and compositions for genetically modifying a cell are provided.

The present invention relates to a guide RNA (gRNA) suitable for CRISPR-mediated oligonucleotide binding and/or editing comprising at least one hairpin that does not interact with a Cas enzyme wherein said hairpin forms a locked secondary structure.

A method for detecting bi-allelic gene edited cells based on CRISPR/Cas12a technology is provided. The method provides crRNA, a nucleotide sequence of which that binds target is SEQ ID NO: 6. The crRNA is transcribed from a transcription template in vitro; the transcription template is a product obtained by annealing a single stranded DNA molecule shown in SEQ ID NO: 10 and a single stranded DNA molecule shown in SEQ ID NO: 13. The method provides crRNA of CD71 gene and constructs a CRISPR/Cas12a system using the same for CRISPR/Cas12a detection, achieving the screening for CRISPR/Cas9 induced CD71 gene bi-allelic gene edited cells. This method is simple, easy, fast, and cost-effective for screening a large number of mutant cells.

The present invention is directed to, inter alia, composition and methods for genome editing. Specifically, a non-naturally occurring OMNI-50 nuclease variant having a wild-type OMNI-50 protein sequence (SEQ ID NO: 1) comprising an amino acid substitution in at least one of the following positions: R61, Y437, R478, A493, Y545, G606, K688, L690, E695, L718, R788, Q803, L805, L844, V981, K965, and K1036.

The disclosure provides Cas12i polypeptides, fusion proteins comprising such Cas12i polypeptides, CRISPR-Cas12i systems comprising such Cas12i polypeptides or fusion proteins, and methods of using the same.