9 results about "Tert gene" patented technology

A DNA immortalization construct includes a first nucleic acid sequence derived from the 5′ end of human CDKN2A gene exon 2. An EF1α promoter sequence is connected to the first sequence. A first LoxP sequence is connected to the EF1α promoter sequence. A human TERT gene sequence is connected to the first LoxP sequence. A second LoxP sequence is connected to the human TERT gene sequence. An internal ribosomal entrance site (IRES) sequence is connected to the second LoxP sequence. An antibiotic selection gene sequence is connected to the IRES sequence. An SV40 poly-A signal sequence is connected to the antibiotic selection gene sequence. A second nucleic acid sequence is connected to the SV40 poly-A signal sequence and derived from 3′ end of human CDKN2A gene exon 2.

The invention discloses a microdroplet type digital PCR (Polymerase Chain Reaction) premixed solution and application thereof in detecting a high-GC target, and belongs to the technical field of gene detection. The microdroplet type digital PCR premixed solution comprises 3 to 5 mM of MgCl2, 60 to 80 mM of KCl, 10 to 30 mM of Tris-HCl with the pH value of 8.3, 150 to 200 nM of dNTP, 0.1 to 0.2 U / l of hot start Taq enzyme, glycerol with the volume fraction of 0.25 to 1 percent, betaine with the volume fraction of 0.25 to 0.75 M, EDTA (Ethylene Diamine Tetraacetic Acid) with the volume fraction of 0.25 to 0.75 mM and formamide with the volume fraction of 0.5 to 2 percent. The premixed liquid has the advantages of good uniformity, high thermal stability, accurate amplification and good Taq enzyme activity maintenance, is suitable for a probe method, provides an efficient universal reagent for a droplet type digital PCR instrument, and realizes high-sensitivity detection of C228T mutation in a TERT gene high GC content (more than 70%) region.

The invention discloses siRNA of a targeted FGF2 / TERT gene, a complex co-delivery system and application of the siRNA. According to the siRNA of the targeted FGF2 / TERT gene and the complex co-delivery system adopting the siRNA, the gene expression of FGF2 or TERT in cells can be remarkably inhibited; the delivery system disclosed by the invention can be used as an FGF2 or TERT inhibitor, remarkably overcomes chemotherapy drug resistance, can also enhance the sensitivity of tumor cells to chemical drugs, has an important clinical application value in the aspect of improving the treatment effect of tumors such as breast cancer and neuroblastoma, and has a great application prospect.

The application belongs to the technical field of fluorescent quantitative PCR, and provides an enzyme digestion-fluorescent quantitative PCR detection method. The method comprises the following steps: step one, configuring a reaction mixture, wherein the reaction mixture comprises amplification reagents, a restriction endonuclease, Taq DNA polymerase, UDG enzyme, primers and a probe; and step two, setting a reaction program and a reaction system. Non-target templates in a qPCR reaction system are degraded by enzyme digestion of a restriction endonuclease, thereby reducing non-specific amplification; the proportion of target templates is increased, thereby avoiding the interference of non-target templates and improving detection sensitivity and detection stability. In particular, the method can effectively detect high GC content templates, such as TERT gene promoters or CpG island methylation.

Disclosed in the present invention is the use of a TERT gene in the detection of residual iPSCs, which can effectively detect the presence of residual iPSCs in differentiated cells derived from hiPSCs, such as iNK cells.

The invention provides an application of joint detection of methylation level and protein concentration in early warning of hepatitis B virus related hepatocellular carcinoma, which adopts joint detection of methylation level of serum TERT gene promoter and TERT protein concentration and is applied to early warning of hepatitis B virus related hepatocellular carcinoma. According to the invention, a method for detecting the methylation level of a TERT gene promoter in free DNA of serum and a method for detecting the protein concentration of TERT in serum are utilized, and the two detection methods are combined into a scheme as a clinical prediction target, so that early warning of hepatitis B virus related hepatocellular carcinoma is realized.

A method for extending the replicative lifespan of T cells by activating the TERT gene is provided. Particularly, the method involves the epigenetic modification of the TERT promoter using a CRISPR / dCas9-based epigenetic modifier, followed by the reactivation of the modified TERT promoter using a Cas9-mediated transcriptional activator.

The present invention relates to the field of cancer. More specifically, the present invention provides methods and compositions related to certain mutations in thyroid cancer. The present inventors hypothesized that SNP rs2853669C>T, by affecting the TERT promoter activities, could differentiate the disease aggressiveness risk associated with BRAF V600E and TERT promoter mutations in PTC and therefore refine their prognostic precision. The combination of these genetic variants in BRAF and TERT genes represents a simple but effective genetic risk prognostication strategy for PTC.

The present invention provides a digital PCR detection reagent and application for BRAF, KRAS, and TERT gene mutations. Specifically, preferred primer pairs, amplification methods, nucleic acid probes, and detection systems targeting sequences containing BRAF, KRAS, and TERT gene mutations are provided. Furthermore, a kit for detecting BRAF, KRAS, and TERT gene mutations is provided. Through optimized primer pairs and probes, as well as corresponding reaction conditions, the present invention can detect BRAF, KRAS, and TERT gene mutations with high sensitivity and specificity in different samples.