TOOLGEN INCORPORATED

The present disclosure relates to targeted genome editing in eukaryotic cells or organisms. More particularly, the present disclosure provides for Cas9/RNA complexes that may induce modifications in target endogenous nucleic acid sequences in nucleuses of eukaryotic cells. The Cas9/RNA complex may comprise a recombinant Cas9 protein including a nuclear localization signal (NLS) and a guide RNA including a crRNA and a tracrRNA. The Cas9/RNA complex may be a combination of the recombinant Cas9 protein and the guide RNA. The guide RNA may be transcribed in vitro or synthesized chemically. The target endogenous nucleic acid sequence may include a portion complementary to the crRNA of the guide RNA.

The present application relates to a method of predicting possible off-targets in a gene editing process (for example, genome editing process) using a gene editing system.

The present disclosure relates to targeted genome editing in eukaryotic cells or organisms. More particularly, the present disclosure provides for compositions and methods that may induce modifications in target endogenous nucleic acid sequences in nucleuses of eukaryotic cells. For example, disclosed herein is a method of producing an engineered eukaryotic cell. In some embodiments, the method may comprise preparing a Cas9 protein, preparing a sgRNA, and preparing a cell-free buffer. The method may further comprise disposing the Cas9 protein and sgRNA in the cell-free buffer to provide for a transfection mixture and transfecting the transfection mixture into a eukaryotic cell, wherein a Cas9/sgRNA complex formed by the Cas9 protein and the sgRNA induces a modification of a target endogenous DNA sequence in the nucleus of the eukaryotic cell to provide for an engineered eukaryotic cell.

The present invention relates to an artificially engineered CRISPR/Cas9 system. More particularly, the present invention relates to an artificially engineered CRISPR enzyme having enhanced target specificity and a use of an artificially engineered CRISPR/Cas9 system including the same enzyme in genome and/or epigenome manipulation or modification, genome targeting, genome editing, and in vitro diagnosis, etc.

A method for producing a potato plant with suppressed browning may use CRISPR/Cas9 system. In a method of producing a genome-edited potato plant with suppressed browning, a potato protoplast is transfected with a ribonucleoprotein complex containing sgRNA which targets StPPO2 ( Solanum tuberosum Polyphenol Oxidase 2) gene derived from potato. Since the method involves no insertion of a foreign gene and includes only tiny mutation that is hardly distinguishable from natural mutation, it is expected that, unlike genetically modified organism (GMO) crops which require a huge amount of cost and time for evaluating the safety and negative environmental impacts, a great deal of cost and time can be saved in the present invention.

The present invention relates to an artificially engineered CRISPR/Cas9 system. More particularly, the present invention relates to an artificially engineered CRISPR enzyme having enhanced target specificity and a use of an artificially engineered CRISPR/Cas9 system including the same enzyme in genome and/or epigenome manipulation or modification, genome targeting, genome editing, and in vitro diagnosis, etc.

The present application relates to a method for predicting an off-target which can occur in the process of editing a genome by using a prime editing system.

The present invention relates to an artificially manipulated SC function-controlling factor for SC function control and/or the treatment or alleviation of a disease due to an SC function disorder, and to a use thereof. More specifically, the present invention relates to a system capable of performing artificial SC function control and/or treating or alleviating a disease due to an SC function disorder, the system comprising: an artificially manipulated SC function-controlling factor for SC function control and/or the treatment or alleviation of a disease due to an SC function disorder; and/or a composition for treating or alleviating a disease due to an SC function disorder. In a specific aspect, the present invention relates to an SC function-controlling system by an SC function-controlling factor, such as artificially manipulated PMP22, and/or an expression product thereof.

The present disclosure relates to targeted genome editing in eukaryotic cells or organisms. More particularly, the present disclosure provides for compositions and methods that may induce modifications in target endogenous nucleic acid sequences in nucleuses of eukaryotic cells. For example, methods disclosed herein may comprise preparing a Cas9/RNA complex, wherein the Cas9/RNA complex comprises a Cas9 protein and a guide RNA. Methods may further comprise introducing the Cas9/RNA complex into a non-human embryo, wherein the Cas9/RNA complex induces a modification at a target endogenous nucleic acid of the non-human embryo to provide for a genome modified embryo. Methods may further comprise transferring the genome modified embryo into a foster mother and allowing the foster mother to produce a F0 animal having the modification at the target endogenous nucleic acid.

The present disclosure discloses a SpCas9 variant. The SpCas9 variant is characterized by the ability to recognize a PAM sequence different from that of a wild-type SpCas9 protein.