Provided herein is an expression vector comprising a coding nucleic acid sequence encoding a polypeptide of interest, wherein the polypeptide of interest is not constitutively expressed from the vector; and a guide-binding sequence located upstream of the coding sequence, wherein the guide-binding sequence comprises a sequence complementary to a nucleic acid guide; wherein binding of a nucleic acid guide to the guide-binding sequence directs a mutation in a nucleic acid sequence of the vector resulting in expression of the polypeptide of interest. Also provided are a combination comprising the expression vector and a nucleic acid guide, a cell comprising the expression vector and/or nucleic acid guide and associated medical methods and uses.

The present invention relates to an adenine base editor (ABE), and components thereof. The present invention also relates to a complex comprising an adenine base editor (ABE) and a guide RNA in a functionally associated form. The present invention further relates to a nucleic acid molecule encoding the ABE/guide RNA, an expression construct or a vector comprising a nucleic acid sequence encoding the adenine base editor and/or the nucleic acid sequence encoding the guide RNA. The present invention further relates to a cell comprising an adenine base editor (ABE) and a method of adenine base editing of a target site in a genome of interest in at least one cell of a prokaryotic organism, including bacterial and archaeal organisms, or eukaryotic organism. Besides that, the present invention relates to various methods, kits and uses associated with the ABEs provided.

The present invention provides recombineering-editing systems using CRISPR and recombination enzymes as well as methods, vectors, nucleic acid compositions, and kits thereof. The methods and systems provide means for altering target DNA, including genomic DNA in a host cell. Specifically, the invention provides systems and compositions utilizing aptamers to bind components of the recombination-editing system such as a CAS protein or guide RNA to enhance targeting of genomic DNA in cells.

The present invention provides methods and compositions for treating cancers, as well as methods for increasing an immune response against a tumor, in a subject in need thereof by decreasing the expression and/or activity of Stub1 in an immune cell of the subject.

The present disclosure provides compositions and methods for reducing body fat in a human male subject by mutating or reducing the expression of the Adgrg6 gene in one or more cells of the human male subject.

A method of inducing SNP mutations in mesenchymal stem cells (MSCs), targeting the most frequent SNP mutations of the TSC2 gene, TSC2.1864C>T (p.Arg622Trp), TSC2.1832 G>A (p.Arg611Glu), and TSC2.5024 C>T (p.Pro1675Leu) using delivery methods for CRISPR components, is described. A high editing efficiency (up to 85%) for inducing TSC2 SNP mutations in MSCs using lipofectamine-based transfection was achieved. Overall, the high editing efficiency of some TSC2 mutations enables the induction and reversal of mutations in primary hMSCs without requiring the resource-consuming derivation of cell lines that are frequently distinct from their primary counterparts.

Disclosed herein are an engineered immune cell with CD7 gene knock-out and use thereof. According to the present invention, an sgRNA specifically targeting CD7 gene is designed and synthesized, which can accurately target CD7 gene to achieve gene knock-out with high knock-out efficiency. The provided sgRNA can be used for preparing a CD7-targeting engineered immune cell, and can be further used for preparing a CD7-targeting universal CAR-T cell

RNA-targeting Cas12a2 complex allows for rationale design of Cas12a2 into a versatile enzyme capable of non-specifically degrading distinct types of nucleic acid targets depending on mutations of the active site residues and residues that stabilize bound targets. These mutations allow for tuning of output signal associated with RNA detection. By mutating specific residues, indiscriminate single-stranded RNase and DNase and double-stranded DNase activity can be modified to only cleave single-stranded DNA and single-stranded RNA, or only single-stranded DNA. This allows for diagnostic tools which can provide a detection. Residues involved in binding the non-self vs. self-recognition signal (PFS) can also be modified so larger subsets of nucleic acid targets can be recognized.

The invention provides methods for enriching nucleic acid target sequences from a sample, for example, from a biological sample or from a nucleic acid library.

Disclosed is a method of genetically recombining lactic acid bacteria and gram-positive bacteria using the CRISPR/Cas system, more specifically, to a method of genetically recombining lactic acid bacteria and gram-positive bacteria that is capable of recombining efficiently lactic acid bacteria and gram-positive bacteria that are difficult to recombine by increasing the efficiency of the RNP recombination system using Cas proteins. However, there is a problem in which it is difficult to recombine genes of Gram-positive bacteria and lactic acid bacteria even using the CRISPR/Cas system due to the cell wall structure thereof. On the other hand, it was found that genes of lactic acid bacteria and gram-positive bacteria that are difficult to recombine can be recombined with high efficiency by using recombinases in combination with a phosphorothioated donor DNA in an RNP recombination system using Cas protein.