13 results about "Tert gene" patented technology

The invention provides a kit for detecting human BRAF gene V600E and TERT gene promoter mutation, and belongs to the technical field of biology. The kit comprises a primer, a probe, a 5 * PCR (Polymerase Chain Reaction)-Enhancer, a 10 * PCR Buffer, dNTP (Deoxyribonucleoside Triphosphate) Mix, MalTaq DNAPlyase, magnesium ions, DEPC (Diethylpyrocarbonate) water, a positive control and a negative control. According to the present invention, the ARMS-PCR detection technology is adopted to detect the specific gene mutation, and the target mutation site is identified by designing the specific primer and the probe, such that the detection specificity and the detection sensitivity are improved; by combining with a Taqman probe technology, the fluorescent signal change in the PCR amplification process can be monitored in real time, so that whether mutation of a BRAF V600E site, a TERT C228T site and a TERT C250T site exists or not is accurately judged; according to the method, pre-amplification of 15 cycles is adopted, then quantitative amplification of 30 cycles is adopted, and when the initial amount of the target DNA is very low, the copy number of the target DNA can be increased, so that the detection accuracy and sensitivity are further improved.

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 invention provides a BRAF, KRAS and TERT gene mutation digital PCR detection reagent and application thereof. Specifically, the invention provides an optimized primer pair, an amplification method, a nucleic acid probe and a detection system aiming at sequences where BRAF, KRAS and TERT gene mutations are located, and further provides a kit for detecting the BRAF, KRAS and TERT gene mutations. According to the invention, the BRAF, KRAS and TERT gene mutation can be detected with high sensitivity and strong specificity for different samples by optimizing the primer pair, the probe and corresponding reaction conditions.

The invention discloses application of a hypoxia inducible factor-1 alpha subunit activator in embryo telomere length recovery, and belongs to the technical field of biological medicine. According to the present invention, embryos at different stages are subjected to single cell sequencing, a Tert gene single cell regulation network is constructed, key transcription factors affecting the specificity of the embryos at the early embryo development stage are screened, and a series of experiments verify that the Hif1alpha achieves the function through HRE in the region 2 of the Tert promoter region, has the transcription regulation effect at the upstream of the Tert, and can be used as a transcription regulation and control gene. Therefore, the telomere length of the embryo is influenced. Therefore, when the fertilized ovum is subjected to in-vitro culture, a hypoxia inducible factor-1alpha subunit activator is added in different stages of embryo culture, Hif1alpha expression is activated, Tert gene expression level is improved, telomere extension of the embryo is realized, and the telomere length of the embryo subjected to in-vitro culture is equivalent to that of an embryo obtained by natural pregnancy.

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 invention relates to an exosome containing TERT mRNA and a preparation method and application thereof, and belongs to the technical field of gene engineering. The exosome is loaded with mRNA (messenger ribonucleic acid) of a TERT (telomerase reverse transcriptase) gene, and the mRNA of the TERT gene is generated by cells over-expressing the TERT gene; the cells for overexpressing the TERT gene are human mesenchymal stem cells for overexpressing the TERT gene. According to the invention, a TERT gene is overexpressed in human mesenchymal stem cells through a genetic engineering technical means, and the exosome is obtained through the cells. Experiments prove that the exosome disclosed by the invention can improve the expression level of related telomere genes and can prolong the length of systemic telomeres. Therefore, the exosome disclosed by the invention has the effect of prolonging systemic telomeres.

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.