43 results about "EIF4E" patented technology

New RNA cap analogs are disclosed containing one or more phosphorothioates groups. The analogs also contain modifications at the 2′-O position of 7-methylguanosine that prevent them from being incorporated in the reverse orientation during in vitro synthesis of mRNA and that hence are “anti-reverse cap analogs” (ARCAs). The ARCA modification ensures that the S atom is precisely positioned within the active sites of cap-binding proteins in both the translational and decapping machinery. The new S-ARCA analogs are resistant to in vivo decapping enzymes. Some S-ARCAs have a higher affinity for eIF4E than the corresponding analogs not containing a phosphorothioate group. When mRNAs containing the various S-ARCAs are introduced into cultured cells, some are translated as much as five-fold more efficiently than mRNAs synthesized with the conventional analog m7GpppG.

The invention provides the components of in vivo and in vitro systems and methods which use them to study the effects of altered expression of a gene activity, such as the human akt, bcl-2, eIF4E or PTEN activities, on the descendants of stem cells that have been engineered to give rise to hematopoietic tumorigenic or tumor cells, such as lymphomas, with a high frequency. The present invention provides vectors, cells and mammals, and methods which in part depend on such products, useful for understanding tumorigenesis and its treatments, and in particular, for identifying and studying inhibitors and activators associated with tumor cell growth and growth inhibition, cell death through apoptotic pathways, and changes in apoptotic pathway components that affect drug sensitivity and resistance in tumorigenic cells. Methods for identifying molecular targets for drug screening, identifying interacting gene activities, for identifying therapeutic treatments and for identifying candidates for new therapeutic treatments are provided.

The present invention relates to methods of imparting virus resistance to plants. In one aspect, this method involves silencing a gene encoding a translation initiation factor eIF4E in the plant. In another aspect, this method involves overexpressing a heterologous translation initation factor eIF4E in a plant. The present invention further relates to a genetic construct containing a nucleic acid molecule encoding a heterologous translation initiation factor eIF4E, as well as to an expression system containing the genetic construct and a host cell transformed with the genetic construct. The present invention also relates to transgenic plants, seeds, and plant parts transformed with the genetic construct. The present invention also relates to an isolated nucleic acid molecule encoding a mutant translation initiation factor eIF4E that is effective in imparting virus resistance in plants. The present invention also relates to a mutant translation initiation factor eIF4E and a method for making the mutant.

Provided are methods and compositions for inhibiting eukaryotic translation initiation factor eIF4E. Such methods and compositions may be used alone or in conjunction with other therapies, such as gene therapies, for inhibiting cell proliferation and / or treating cancer.

Oligomeric compounds, compositions and methods are provided for modulating the expression of eIF4E. The antisense compounds may be single- or double-stranded and are targeted to nucleic acid encoding eIF4E. Methods of using these compounds for modulation of eIF4E expression and for diagnosis and treatment of diseases and conditions associated with expression of eIF4E are provided.

The invention discloses two site-specific dominant ASM markers which are directly related with the identification of a wild type and a mutant of eIF4E.a of Chinese cabbage. One marker is that: a marker which is related with the detection of a wild type site is named ASM-4E.a-s and is shown as SEQ ID No.1; and a marker which is related with the detection of the corresponding mutation site is namedASM-4e.a-s and is shown as SEQ ID No.3. The other marker is that: a marker which is related with the detection of a wild type site is named ASM-4E.a-lc and is shown as SEQ ID No.2; and a marker whichis related with the detection of the corresponding mutation site is named ASM-4e.a-1c and is shown as SEQ ID No.4. The invention also discloses a primer for identifying the two specific dominant molecular markers. The primer is shown as SEQ ID NO.5, SEQ ID NO.6 and SEQ ID NO.7. The markers and the primer can be taken as molecular markers and applied to the identification of germplasm resources ofthe Chinese cabbage and the aided selection of breeding; and a germplasm material or a transformation offspring containing an eIF4E.a mutation type in 06-247 or Guan291 is selected.

Prepared is an extract composition having an improved protein synthetic activity in a cell-free protein synthesis system using a mammalian cultured cell extract. An eukaryotic translation initiation factor and / or translational regulator are added to a cell-free protein synthesis system comprising an extract prepared from cultured mammalian cells and a template mRNA. These factors are one or more selected from the group consisting of eukaryotic translation initiation factors 4E (eIF4E), 2 (eIF2) and 2B (eIF2B), and eukaryotic translational regulator p97.

The invention provides a process for preparing capped mRNAs from an RNA mixture, e.g. whole RNA isolated from a cell or tissue extract, that includes combining in a reaction mixture RNA comprising capped mRNA with a separable affinity matrix having high-affinity eIF4E bound thereto, under conditions sufficient for binding to occur between the high-affinity eIF4E and the capped mRNA, whereby capped mRNA is bound to the affinity matrix, separating the affinity matrix from the reaction mixture, then separating the capped mRNA from the affinity matrix. High affinity eIF4E mutants previously described are employed in the process as well as a novel mutant disclosed and claimed herein. The mRNA preparation process is based on isolation of 5′-capped mRNA. The mRNA molecules thus isolated have intact sequences encoding the NH2-terminal ends of the proteins they encode, unlike those isolated by prior methods. In addition, use of the method isolates mRNA sequences not isolatable by prior methods that relied on binding to polyadenylated 3′-end sequences.

The present invention provides compounds to disrupt the eIF4E-eIF4G interaction and pharmaceutically acceptable salts of such compounds. Generally, the compounds are cell-penetrating peptides which bind mammalian initiation factor eIF4E (CPP-eIF4E), wherein the peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOS: 7, 9-11, 12-26, 27-44 and 45-51. More preferably, the amino acid sequence comprises at least 9 to about 40 amino acids and the mammalian initiation factor eIF4E is human initiation factor eIF4E.

Prepared is an extract composition having an improved protein synthetic activity in a cell-free protein synthesis system using a mammalian cultured cell extract. An eukaryotic translation initiation factor and / or translational regulator are added to a cell-free protein synthesis system comprising an extract prepared from cultured mammalian cells and a template mRNA. These factors are one or more selected from the group consisting of eukaryotic translation initiation factors 4E (eIF4E), 2 (eIF2) and 2B (eIF2B), and eukaryotic translational regulator p97.

Compounds, compositions and methods are provided for modulating the expression of eIF4E-BP1. The compositions comprise oligonucleotides, targeted to nucleic acid encoding eIF4E-BP1. Methods of using these compounds for modulation of eIF4E-BP1 expression and for diagnosis and treatment of diseases and conditions associated with expression of eIF4E-BP1 are provided.

New RNA cap analogs are disclosed containing one or more phosphorothioates groups. The analogs also contain modifications at the 2′-O position of 7-methylguanosine that prevent them from being incorporated in the reverse orientation during in vitro synthesis of mRNA and that hence are “anti-reverse cap analogs” (ARCAs). The ARCA modification ensures that the S atom is precisely positioned within the active sites of cap-binding proteins in both the translational and decapping machinery. The new S-ARCA analogs are resistant to in vivo decapping enzymes. Some S-ARCAs have a higher affinity for eIF4E than the corresponding analogs not containing a phosphorothioate group. When mRNAs containing the various S-ARCAs are introduced into cultured cells, some are translated as much as five-fold more efficiently than mRNAs synthesized with the conventional analog m7GpppG.

The invention discloses a specific co-dominance molecular marker for large-fragment deletion mutation of an eIF4E-1 site of tobacco and application of the molecular marker. A marker relevant with thedetection of a wild site is named as ASM-W and is represented by SEQ ID No.1, and the fragment size is 763bp; and a marker relevant with the detection of a corresponding base deletion mutation site isnamed as ASM-m and is represented by SEQ ID No.2, and the fragment size is 572bp. The invention further discloses a primer for identifying the specific co-dominance molecular marker for the large-fragment deletion mutation, and the sequences of the primer are represented by SEQ ID NO.3, 4 and 5. By utilizing the markers and the primer, the genotype of backcross transformation descendants of PVY resistance donor parents can be accurately selected. The markers and the primer can be applied to the germplasm resource identification and assist breeding of the tobacco and the selection of germplasmmaterials or transformed descendants in the PVY resistance mutation type.

The invention provides a method of purifying human eIF-4E protein and amino acid sequence variants thereof having altered binding affinity for capped RNA. Using the described purification, amino acid sequence variants can readily be expressed, purified and tested. Both lowered and enhanced binding affinity variants are useful for modifying protein expression levels in vivo and in vitro.

The invention provides application of homoharringtonine-type compounds in antitumor drug preparation, especially aiming at p-eIF4E expressed positive tumors. Phosphorylated eIF4E(p-eIF4E), which is separated and identified from tumor cells by biotin-labeled HHT (homoharringtonine), is one of important anti-tumor target molecules of the homoharringtonine-type compounds; studies show that the p-eIF4E is in an extremely high expression state in some tumors, and the homoharringtonine-type compounds have remarkable inhibition effect on the p-eIF4E expressed positive tumors. The homoharringtonine-type compounds can be applied to preparation of drugs for treating the p-eIF4E expressed positive tumors.

The invention relates to an RNAi (RNA interference) vector construction method and an application thereof in watermelon genetic transformation, relates to the technical field of plant virus control and belongs to the application technology in biological and modern agriculture technique fields. The mediated virus infection function is destroyed by acting on host factors and thus watermelons obtain antiviral activity, which has not been reported yet at present. An RNAi vector is constructed and genetically transformed to watermelons, the constructionmethod is characterized in that watermelon eIF4E and a homological isomer eIF(iso)4E gene thereof are cloned and connected, 4E-iso sense and antisense fragments are obtained and connected to two sides of an intron sequence, and a 4E-iso hairpin structure fragment is obtained; the 4E-iso sense fragment, the 4E-iso antisense fragment and the hairpin structure fragment are inserted into a pCAMBIA1301 vector through T4 connection and homologous recombination, and the RNAi vector is obtained; target gene fragments are transformed into watermelon cotyledon explants through agrobacterium tumefaciens-medicated transformation, and transgenic watermelons with the target gene fragments are obtained through screening cultivation. The method is mainly used for providing a novel material and a novel technology for watermelon virus and disease control.

Disclosed herein is a method of inhibiting T Follicular Helper (TFH) cell-mediated differentiation and / or activation in a subject. This method involves administering to a subject in need of treatment for an autoimmune disorder a eukaryotic translation initiation factor 4E (eIF4E) inhibitor to inhibit TFH cell-mediated differentiation and / or activation in the subject. Also disclosed is a method of inhibiting T Follicular Helper (THF) cell differentiation or TFH cell activity.

The invention discloses a preparation method of an anti-tobacco potato Y virus vaccine. The method includes the following steps that 1, pCaPVX440-eIF4E-6 plasmids are prepared; 2, the pCaPVX440-eIF4E-6 is transferred to agrobacterium to prepare the anti-tobacco potato Y virus vaccine. The anti-tobacco potato Y virus vaccine prepared by means of the preparation method is suitable for prevention and control of tobacco PVY viruses and has an important practical value for prevention and control of diseases caused by the PVY viruses in the tobacco leaf production process. Compared with the prior art, the virus vaccine is good in specificity, good in safety, simple, convenient and quick to operate and low in cost.

The invention discloses two site-specific dominant ASM markers which are directly related with the identification of a wild type and a mutant of eIF4E.a of Chinese cabbage. One marker is that: a marker which is related with the detection of a wild type site is named ASM-4E.a-s and is shown as SEQ ID No.1; and a marker which is related with the detection of the corresponding mutation site is namedASM-4e.a-s and is shown as SEQ ID No.3. The other marker is that: a marker which is related with the detection of a wild type site is named ASM-4E.a-lc and is shown as SEQ ID No.2; and a marker whichis related with the detection of the corresponding mutation site is named ASM-4e.a-1c and is shown as SEQ ID No.4. The invention also discloses a primer for identifying the two specific dominant molecular markers. The primer is shown as SEQ ID NO.5, SEQ ID NO.6 and SEQ ID NO.7. The markers and the primer can be taken as molecular markers and applied to the identification of germplasm resources ofthe Chinese cabbage and the aided selection of breeding; and a germplasm material or a transformation offspring containing an eIF4E.a mutation type in 06-247 or Guan291 is selected.

The invention discloses a silkworm PABP (polyadenylic acid bonding protein) bonding protein interacting factor gene BmPaip1. The nucleotide sequence of the gene BmPaip1 is shown as SEQIDNO.3, the open reading frame of the gene BmPaip1 is 1194bp long, composed of eight exons and codes 398 amino acids, theoretical protein molecular weight is 44kDa, and isoelectric point pI is equal to 5.02; and SMART online software analysis shows that the bits from 100 to 398 in the amino acid sequence of the gene BmPaip1 of a silkworm are highly homologous with an eIF4G protein of an eucaryon, and expression detection in insect cells shows that expression level of a reporter gene LUC (luciferase) can be obviously improved in cells genetically modified by the gene BmPaip1 of the silkworm, so that the gene BmPaip1 of the silkworm can be combined with eIF4E and eIF4A to form a translation initiation complex and can be used for improving translation efficiency of an exogenous gene.

A peptide that binds to elongation initiation factor 4E (eIF4E) comprising the amino acid sequence CEX1GX2X3X4X5C (SEQ ID NO: 1), where X1 is an amino acid selected from the group consisting of threonine (T), methionine (M) or leucine (L), X2 and X3 is an amino acid selected from the group consisting of phenylalanine (F), modified phenylalanine and tyrosine (Y), X4 and X5 is any amino acid, wherein the two cysteine residues are joined by a disulphide bond. Further, pharmaceutical compositions and uses of the peptide, and pharmaceutical compositions are provided.

The invention provides a method of purifying human eIF-4E protein and amino acid sequence variants thereof having altered binding affinity for capped RNA. Using the described purification, amino acid sequence variants can readily be expressed, purified and tested. Both lowered and enhanced binding affinity variants are useful for modifying protein expression levels in vivo and in vitro.