Invented by Tian Xu, Feng Qian, Sean LANDRETTE, Yale University

Transposons are DNA sequences that can move from one location to another within a genome. They are often called “jumping genes” because of their ability to move around. Transposons can have a significant impact on the genome, as they can disrupt genes, cause mutations, and even create new genes. In recent years, there has been a growing interest in the market for compositions of transposons and methods for using them. The market for compositions of transposons and methods for using them is driven by several factors. One of the main drivers is the increasing demand for gene therapy. Gene therapy is a promising field that aims to treat genetic disorders by introducing new genes into the body. Transposons can be used as a delivery system for these new genes, making them an attractive option for gene therapy. Another driver of the market is the growing interest in genome editing. Genome editing is a technique that allows scientists to make precise changes to the DNA sequence of an organism. Transposons can be used to deliver the necessary enzymes for genome editing, making them a valuable tool for this field. The market for compositions of transposons and methods for using them is also driven by the increasing demand for agricultural biotechnology. Transposons can be used to introduce new traits into crops, such as resistance to pests or drought. This can help to increase crop yields and improve food security. There are several companies that are active in the market for compositions of transposons and methods for using them. One of the leading companies in this field is Transposagen Biopharmaceuticals. Transposagen offers a range of products and services related to transposon technology, including custom transposon design, transposon-mediated gene delivery, and genome editing. Another company that is active in this field is Sigma-Aldrich. Sigma-Aldrich offers a range of transposon-related products, including transposon vectors and transposon-mediated gene delivery systems. In conclusion, the market for compositions of transposons and methods for using them is a growing and promising field. The increasing demand for gene therapy, genome editing, and agricultural biotechnology is driving the growth of this market. As the field continues to develop, we can expect to see new and innovative uses for transposons emerge.

The Yale University invention works as follows

The present invention consists of compositions and techniques for using transposons. In one aspect, methods that can be used to identify negatively selected genes during an insertional mutation screen are described. Compositions that reduce proliferation of tumor cells expressing oncogenic RAS in another aspect include an activator for the WNT pathway. “Pharmaceutical compositions are disclosed for reducing the proliferation of tumor cells by administering an active amount of an activated WNT pathway in the tumor cells.

Background for Compositions of transposons and methods for using them

Recent advances in sequencing technology have enabled the identification of specific mutants in specific tumors. This opens the door to developing targeted therapies for specific tumors. Functional genomics is a powerful tool for identifying the drivers behind human disease and biological processes. CRISPR Cas9 and shRNA library screening tools are effective in knocking out or knocking down protein-coding gene. Some cancers, such as HER2-amplified breast cancer or acute promyelocytic lymphoma, can be treated by targeting specific oncogenic alterations in tumor cells. Directly targeting oncogenic pathways or alterations has proved difficult for some common mutations, such as activating RAS, and losing TP53. Many diseases and biological traits are caused by abnormal gene activation or overexpression. It is therefore highly desirable to use forward genetic screens to examine the human genome to identify synthetic lethal interactions within tumor cells that have oncogenic mutations. “While loss-of function screens on cancer cell lines using shRNA libraries were successfully used to identify synthetic lethal target genes, genome-wide gains-of-function screenings for negatively selected gene are lacking.

Therefore there is a need in the field of science for improved methods that can identify genes with a negative selection, particularly in the case where oncogenic mutations are common and lead to cancer.

As described below, this invention includes compositions and methods for identifying therapeutic target and pathways specific to tumor cells by negative selecting genes on an insertional mutation screen.

The invention includes a way to identify negatively selected genes from an insertional mutation screen. This involves introducing a piggyBac in cells of interest, exposing a part of these cells to selective pressure in order to induce the expression of the piggyBac; comparing the insertion site in genomic DNA between transposed and non-transposed cells; and identifying the genes that have one or more sites.

Another aspect of the invention is a composition that reduces proliferation of tumor cells expressing oncogenic RAS and contains an activator of the WNT pathway.

Yet another feature of the invention is a pharmaceutical formulation containing the composition described herein, and a pharmaceutically accepted carrier.

Yet another aspect of invention is a method for reducing the proliferation of tumors in a patient in need of it, which comprises administering a sufficient amount of a formulation containing an activator of WNT pathway in the tumor cells of that subject and reducing the proliferation of those tumor cells.

Another aspect” of the invention is a method for reducing or improving cancers expressing oncogenic RAS, and/or symptoms associated with it in a patient by administering an activater of a WNT path.

Yet another embodiment of the invention comprises a composition that can be used to treat an oncogenic RAS tumour, the composition containing an activator for a WNT path.

In various embodiments, or in any aspect of this invention described herein, a piggyBac-transposon contains an antibiotic resistance gene that can be induced. In one embodiment the cells of concern are cancer cells. The tumor cells can be at least one from the following: lung, liver, gastrointestinal (colon), pancreatic and skin cells. In another embodiment, propagating transposed cells of concern is included in the step of inducing the transposition. In a further embodiment, the comparison of insertion site comprises the sequencing of the insertion site. In another embodiment, insertion sites can be located in an exon, an intron or a promoter of a gene. In yet another embodiment the genes are present in transposed cells that have not been exposed to selective pressure, but depleted in transposed cell exposed to selective pressure. Another embodiment is that the genes inhibit the growth or survival of cells of interest.

In one embodiment, an activator is a GSK inhibitor. In another embodiment, the activator is selected from the group consisting of 2-Amino-4-(3,4-(methylenedioxy)benzylamino)-6-(3-methoxyphenyl)pyrimidine, LiCl, Kenpaullone and 6-bromoindirubin-30-oxime (BIO). In another embodiment, activator is small molecule WNT pathway agonist.

In another embodiment, an oncogenic RAS can be selected from a group that includes oncogenic HRASs, oncogenic NRAs, and oncogenic KRASs.

In a further embodiment, the composition is further formulated to deliver at least one of the following: a skin, lung, liver or gastrointestinal tumor.

In some embodiments, the composition is used to treat a cancer that has oncogenic Ras expressed in its cells. The composition includes a first agent which is an agonist for one or more WNT pathway members. In some embodiments the composition includes a second agent which is an antagonist of Ras.

In some embodiments, the method for treating cancer characterized by expression of oncogenic Ras within the cancer cells involves administering a composition to a patient with the cancer that contains an effective amount of first agent which is an agonist of at least one member of the WNT pathway. This will treat the cancer. In some embodiments the method includes administering an amount of a second agent which is an antagonist to oncogenic Ras. The effective amount of first or second agent in some embodiments is an amount that inhibits proliferation of cancer cells.

In some embodiments, a first agent is a small molecule agonist of WNT pathway. The group of genes that are selected for this purpose includes LRP6, -catenin and?-catenin; TCF7L1, CSNK1G1, CCNY; PCDH15; GNG7; INO80. SMARCC1, PRKCA and MED13. In some embodiments the first agent is small molecule WNT pathway agonist. In some embodiments, the first agent is selected from the group consisting of a glycogen synthase kinase (GSK) inhibitor, 2-Amino-4-(3,4-(methylenedioxy)benzylamino)-6-(3-methoxyphenyl)pyrimidine, LiCl, Kenpaullone and 6-bromoindirubin-30-oxime (BIO), and pharmaceutically acceptable salts, analogs, and derivatives thereof.

In some embodiments, cancer cells express a RAS that is selected from a group of oncogenic HRASs, oncogenic NRAss and oncogenic Krass.

Definitions

The terms are used in this document to describe the subject matter of the invention. The preferred materials and methods described in this document are those that can be used to test the invention. However, any materials or methods similar or equivalent to the ones described herein could also be used. The following terminology will apply to describing and claiming this invention.

It is important to note that the terms used in this document are not meant to limit the invention.

The articles?a. The articles?a? The words?an? and?a? are used to indicate one or more (i.e. at least one) of grammatical objects of an article. As an example, “an element” is used. “An element” means a single element or multiple elements.

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