32 results about "Insertional mutagenesis" patented technology

A modified P. aeruginosa type III secretion system has been developed that efficiently delivers selected proteins into a host cell. In one example, a functional nuclear Cre Recombinase is injected into embryonic stem (ES) cells and can be used to induce pluripotent stem (iPS) cells. This method of in vitro lineage directed differentiation prevents insertional mutagenesis and provides a route to selected stem cell renewal and cell-based therapies.

The present invention provides hyperactive piggyBac transposons, in particular hyperactive piggyBac transposons from Trichoplusia ni (cabbage looper moth) that transpose at a higher frequency than wildtype. The invention also features integration defective piggyBac transposons. The piggyBac transposons and transposases can be used in gene transfer systems for stably introducing nucleic acids into the DNA of a cell. The gene transfer system can be used in methods, for example, but not limited to, gene therapy, insertional mutagenesis, or gene discovery.

The present invention provides polynucleotide vectors for high expression of heterologous genes. Some vectors further comprise novel transposons and transposases that further improve expression. Further disclosed are vectors that can be used in a gene transfer system for stably introducing nucleic acids into the DNA of a cell. The gene transfer systems can be used in methods, for example, gene expression, bioprocessing, gene therapy, insertional mutagenesis, or gene discovery.

The rice dwarf gene relates to a rice gene, in particular to a rice dwarf gene obtained by T-DNA insertion mutation method. Provided are an osDw gene obtained by a method of T-DNA insertion mutation and its cloning method; an osDw gene RNAi expression vector and its construction method; a genetic transformation method of the osDw gene and its application in genetic transformation. The full length of the gene is 2720 bases, in which ATG at the 33rd to 35th is the start codon, and the TGA sequence at the 2253rd to 2255th is the stop codon. The cloning method is to amplify a 2.7kb nucleic acid fragment by PCR method based on the nucleic acid sequence published on NCBI based on the results of blast analysis; the nucleic acid fragment is recovered and confirmed by sequencing after being connected to the T carrier. It has the function of controlling the plant height of rice. The RNAi vector can be constructed according to its sequence, and dwarf plants can be obtained by transgenic method.

The present invention relates to methods of selecting for highly productive clones of E. coli for the production of plasmid DNA comprising measuring the frequency of IS1 transposon insertional mutagenesis within either the plasmid or genomic DNA of transformed clonal subtypes. An increase in IS1 insertional mutagenesis is correlated with clonal subtypes likely to exhibit a low specific productivity. The PCR-based, genetic selection assays disclosed herein are amenable to high throughput analysis, reducing the time to identify highly productive clones capable of cultivating large quantities of plasmid DNA on an industrial scale.

The present invention provides compositions and methods of using transposons. In one aspect, methods are disclosed that are useful for identifying negatively selected genes in an insertional mutagenesis screen. In another aspect, compositions for reducing proliferation of a tumor cell expressing an oncogenic RAS include an activator of a WNT pathway. Pharmaceutical compositions for reducing proliferation of tumor cells in a subject in need thereof by administering an effective amount of an activator of a WNT pathway to the tumor cells of the subject are also disclosed.

The present invention is directed generally to reduction or inactivation of gene function or gene expression in cells in vitro and in multicellular organisms. The invention encompasses methods for mutating cells using a combination of mutagens, particularly wherein at least one mutagen is an insertional mutagen, to achieve homozygous gene mutation or mutation of multiple genes required cumulatively to achieve a phenotype to create knock-outs, knock-downs, and other modifications in the same cell. The invention is also directed to cells (and libraries thereof) and organisms created by the methods of the invention, including those in which at least one of the genes created by insertional mutagenesis is tagged by means of the insertion sequences thereby allowing identification of the mutated gene(s). The invention is also directed to libraries of mutated cells and their uses. The invention is also directed to methods of identifying mutations with methods of the invention, in cells (and libraries thereof) and organisms, by means of the insertional tag.

The invention discloses a high-efficiency transgene method mediated by a transcriptional activator-like effector protein. A helper plasmid containing the DNA sequence of transcriptional activator-like effector protein and piggyBac transposase DNA sequence was constructed by molecular biology techniques, and transcribed into mRNA in vitro; the mRNA of the helper plasmid was mixed with piggyBac containing exogenous gene by microinjection The DNA of the transposon plasmid is introduced into the fertilized egg of the animal together to obtain a transgenic animal in which the exogenous gene can be stably inherited and expressed; the transgenic positive individual is screened out according to the expression characteristics of the marker gene or the gene; the method of the present invention can make piggyBac transposition Under the action of the TALE-PBase fusion protein, the piggyBac transposon can be efficiently inserted into the genome of the host organism, which can significantly improve the transgenic efficiency mediated by the piggyBac transposon, and provide assistance for obtaining transgenic animals and insertional mutagenesis.