In vivo monitoring of retrovirus-tagged blood stem and progenitor cells is used to check hematopoiesis. Two strategies are used most incessantly: sequencing the locus of retrovirus insertion, termed integration web site evaluation, or retrovirus DNA barcode sequencing.
Of these, integration web site evaluation is at present the one out there method for monitoring clonal swimming pools in sufferers handled with retrovirus-modified blood cells.
A key query is how these two strategies examine in their potential to detect and quantify clonal contributions. In this research, we assessed each strategies concurrently in a clinically related nonhuman primate mannequin of autologous, myeloablative transplantation.
Our knowledge show that each strategies observe ample clones; nonetheless, DNA barcode sequencing is no less than 5-fold extra environment friendly than integration web site evaluation. Using computational simulation to establish the sources of low effectivity, we establish sampling depth as the main issue.
We present that the sampling required for integration web site evaluation to realize minimal protection of the true clonal pool is probably going prohibitive, particularly in circumstances of low gene-modified cell engraftment. We additionally present that early subsampling of various blood cell lineages provides worth to clone monitoring data in phrases of security and hematopoietic biology.
Our evaluation demonstrates DNA barcode sequencing as a helpful information to maximise integration web site evaluation interpretation in gene remedy sufferers.
Integrated DNA methylation evaluation reveals a possible position for ANKRD30B in Williams syndrome.
Williams syndrome (WS) is a uncommon genetic dysfunction, brought on by a microdeletion on the 7q11.23 area. WS reveals a large spectrum of options together with hypersociability, which contrasts with social deficits sometimes related to autism spectrum issues. The phenotypic variability in WS probably entails epigenetic modifications; nonetheless, the character of those occasions stays unclear.
DNA Barcoding in Nonhuman Primates Reveals Important Limitations in Retrovirus Integration Site Analysis.
To higher perceive the position of epigenetics in WS phenotypes, we built-in DNA methylation and gene expression profiles in blood from sufferers with WS and controls.
From these research, 380 differentially methylated positions (DMPs), situated all through the genome, have been recognized. Systems-level evaluation revealed a number of co-methylation modules linked to intermediate phenotypes of WS, with the top-scoring module associated to neurogenesis and growth of the central nervous system.
Description: The p65 (RELA) heterodimer is the most abundant form of NF-KB. This gene is located on 11q13, which consists of 10 exons and spans about 8.1 kb of DNA. In rat sciatic nerves, the expression of the activated p65 subunit of NFKB was high in the nuclei of premyelinating Schwann cells and then progressively declined until it was nearly absent in adults. The transcriptional activity of NF-kappa-B is stimulated upon phosphorylation of its p65 subunit on serine-276 by protein kinase A(PKA). The transcriptional coactivator CBP/p300 associates with NF-kappa-B through 2 sites, an N-terminal domain that interacts with the C-terminal region of the unphosphorylated protein, and a second domain that only interacts with p65 phosphorylated on serine-276.
Description: RELA, also called NFKB3 or NFKB, p65 subunit is part of the NF-kB complex. The complex is inhibited by I-kappa-B proteins, which inactivates NFkB by trapping it in the cytoplasm. The p65 heterodimer is the most abundant form of NF-kB. It is a nonhistone substrate of HDAC3 and that IKBA-dependent nuclear export of the HDAC3-deacetylated RELA replenishes the depleted cytoplasmic pool of latent NFKB-IKBA complexes for subsequent NF-kB responses. Its nucleocytoplasmic redistribution coincided with export of PPARG, and immunoprecipitation analysis indicated that PPARG-RELA association was dependent on the PPARG C-terminal ligand-binding domain. IKK-dependent phosphorylation of RELA on Ser468 enhanced binding of GCN5 to RELA and its ubiquitination.
Description: Transcription factor p65, also known as NFKB3 or NF-kB p65, is a protein that in humans is encoded by the RELA gene. It is mapped to 11q13.1. NFKB is an essential transcription factor complex involved in all types of cellular processes, including cellular metabolism, chemotaxis, etc, and it may play a role in inflammatory conditions of the peripheral nervous system. Phosphorylation and acetylation of NFKB3 are crucial post-translational modifications required for NFKB activation. It has also been shown to modulate immune responses, and activation of NFKB3 is positively associated with multiple types of cancer. In addition to that, NFKB3 antagonizes TNFR1-JNK proliferative signals in epidermis and plays a nonredundant role in restraining epidermal growth.
Description: Proteins encoded by the v-Rel viral oncogene and its cellular homolog, c-Rel, are members of a family of transcription factors that include the two subunits of the transcription factor N B (p50 and p65) and the Drosophila maternal morphogen, dorsal. Both proteins specifically bind to DNA sequences that are the same or slight variations of the 10 bp B sequence in the immunoglobulin light chain enhancer. This same sequence is also present in a number of other cellular and viral enhancers. The DNA binding activity of NF B is activated and NF B is subsequently transported from the cytoplasm to the nucleus in cells exposed to mitogens or growth factors. cDNAs encoding precursors for two distinct proteins of the same size have been described, designated p105 and p100. The p105 precursor contains p50 at its N-terminus and a C-terminal region that when expressed as a separate molecule, designated pdI, binds to p50 and regulates its activity.
Description: Proteins encoded by the v-Rel viral oncogene and its cellular homolog, c-Rel, are members of a family of transcription factors that include the two subunits of the transcription factor N B (p50 and p65) and the Drosophila maternal morphogen, dorsal. Both proteins specifically bind to DNA sequences that are the same or slight variations of the 10 bp B sequence in the immunoglobulin light chain enhancer. This same sequence is also present in a number of other cellular and viral enhancers. The DNA binding activity of NF B is activated and NF B is subsequently transported from the cytoplasm to the nucleus in cells exposed to mitogens or growth factors. cDNAs encoding precursors for two distinct proteins of the same size have been described, designated p105 and p100. The p105 precursor contains p50 at its N-terminus and a C-terminal region that when expressed as a separate molecule, designated pdI, binds to p50 and regulates its activity.
Description: Proteins encoded by the v-Rel viral oncogene and its cellular homolog, c-Rel, are members of a family of transcription factors that include the two subunits of the transcription factor N B (p50 and p65) and the Drosophila maternal morphogen, dorsal. Both proteins specifically bind to DNA sequences that are the same or slight variations of the 10 bp B sequence in the immunoglobulin light chain enhancer. This same sequence is also present in a number of other cellular and viral enhancers. The DNA binding activity of NF B is activated and NF B is subsequently transported from the cytoplasm to the nucleus in cells exposed to mitogens or growth factors. cDNAs encoding precursors for two distinct proteins of the same size have been described, designated p105 and p100. The p105 precursor contains p50 at its N-terminus and a C-terminal region that when expressed as a separate molecule, designated pdI, binds to p50 and regulates its activity.
Description: Proteins encoded by the v-Rel viral oncogene and its cellular homolog, c-Rel, are members of a family of transcription factors that include the two subunits of the transcription factor N B (p50 and p65) and the Drosophila maternal morphogen, dorsal. Both proteins specifically bind to DNA sequences that are the same or slight variations of the 10 bp B sequence in the immunoglobulin light chain enhancer. This same sequence is also present in a number of other cellular and viral enhancers. The DNA binding activity of NF B is activated and NF B is subsequently transported from the cytoplasm to the nucleus in cells exposed to mitogens or growth factors. cDNAs encoding precursors for two distinct proteins of the same size have been described, designated p105 and p100. The p105 precursor contains p50 at its N-terminus and a C-terminal region that when expressed as a separate molecule, designated pdI, binds to p50 and regulates its activity.
Description: Proteins encoded by the v-Rel viral oncogene and its cellular homolog, c-Rel, are members of a family of transcription factors that include the two subunits of the transcription factor N B (p50 and p65) and the Drosophila maternal morphogen, dorsal. Both proteins specifically bind to DNA sequences that are the same or slight variations of the 10 bp B sequence in the immunoglobulin light chain enhancer. This same sequence is also present in a number of other cellular and viral enhancers. The DNA binding activity of NF B is activated and NF B is subsequently transported from the cytoplasm to the nucleus in cells exposed to mitogens or growth factors. cDNAs encoding precursors for two distinct proteins of the same size have been described, designated p105 and p100. The p105 precursor contains p50 at its N-terminus and a C-terminal region that when expressed as a separate molecule, designated pdI, binds to p50 and regulates its activity.
Description: Proteins encoded by the v-Rel viral oncogene and its cellular homolog, c-Rel, are members of a family of transcription factors that include the two subunits of the transcription factor N B (p50 and p65) and the Drosophila maternal morphogen, dorsal. Both proteins specifically bind to DNA sequences that are the same or slight variations of the 10 bp B sequence in the immunoglobulin light chain enhancer. This same sequence is also present in a number of other cellular and viral enhancers. The DNA binding activity of NF B is activated and NF B is subsequently transported from the cytoplasm to the nucleus in cells exposed to mitogens or growth factors. cDNAs encoding precursors for two distinct proteins of the same size have been described, designated p105 and p100. The p105 precursor contains p50 at its N-terminus and a C-terminal region that when expressed as a separate molecule, designated pdI, binds to p50 and regulates its activity.
Description: Transcription factor p65, also known as NFKB3 or NF-kB p65, is a protein that in humans is encoded by the RELA gene. It is mapped to 11q13.1. RELA is an essential transcription factor complex involved in all types of cellular processes, including cellular metabolism, chemotaxis, etc, and it may play a role in inflammatory conditions of the peripheral nervous system. Phosphorylation and acetylation of RELA are crucial post-translational modifications required for NFkB activation. It has also been shown to modulate immune responses, and activation of the protein is positively associated with multiple types of cancer. In addition, RELA antagonizes TNFR1-JNK proliferative signals in epidermis and plays a nonredundant role in restraining epidermal growth.
Description: NF-kappa-B is a pleiotropic transcription factor present in almost all cell types and is the endpoint of a series of signal transduction events that are initiated by a vast array of stimuli related to many biological processes such as inflammation, immunity, differentiation, cell growth, tumorigenesis and apoptosis. NF-kappa-B is a homo- or heterodimeric complex formed by the Rel-like domain-containing proteins RELA/p65, RELB, NFKB1/p105, NFKB1/p50, REL and NFKB2/p52 and the heterodimeric p65-p50 complex appears to be most abundant one. The dimers bind at kappa-B sites in the DNA of their target genes and the individual dimers have distinct preferences for different kappa-B sites that they can bind with distinguishable affinity and specificity. Different dimer combinations act as transcriptional activators or repressors, respectively. NF-kappa-B is controlled by various mechanisms of post-translational modification and subcellular compartmentalization as well as by interactions with other cofactors or corepressors. NF-kappa-B complexes are held in the cytoplasm in an inactive state complexed with members of the NF-kappa-B inhibitor (I-kappa-B) family. In a conventional activation pathway, I-kappa-B is phosphorylated by I-kappa-B kinases (IKKs) in response to different activators, subsequently degraded thus liberating the active NF-kappa-B complex which translocates to the nucleus. NF-kappa-B heterodimeric p65-p50 and p65-c-Rel complexes are transcriptional activators. The NF-kappa-B p65-p65 complex appears to be involved in invasin-mediated activation of IL-8 expression. The inhibitory effect of I-kappa-B upon NF-kappa-B the cytoplasm is exerted primarily through the interaction with p65. p65 shows a weak DNA-binding site which could contribute directly to DNA binding in the NF-kappa-B complex. Associates with chromatin at the NF-kappa-B promoter region via association with DDX1. Essential for cytokine gene expression in T-cells. [UniProt]
Description: NF-kappa-B p105/p50, also called EBP-1 is a protein that in humans is encoded by the NFKB1 gene. By fluorescence in situ hybridization, the gene was assigned to human chromosome 4q24. NF-kappa-B is a pleiotropic transcription factor present in almost all cell types and is the endpoint of a series of signal transduction events that are initiated by a vast array of stimuli related to many biological processes such as inflammation, immunity, differentiation, cell growth, tumorigenesis and apoptosis. NFKB1 appears to have dual functions such as cytoplasmic retention of attached NF-kappa-B proteins by p105 and generation of p50 by a cotranslational processing.
Description: Quantitative sandwich ELISA for measuring Human Nuclear factor-KB p65 (NF-KB p65) in samples from cell culture supernatants, serum, whole blood, plasma and other biological fluids.
ELISA kit for Human Nuclear factor-KB p65 (NF-KB p65)
Description: Quantitative sandwich ELISA for measuring Human Nuclear factor-KB p65 (NF-KB p65) in samples from cell culture supernatants, serum, whole blood, plasma and other biological fluids.
ELISA kit for Human Nuclear factor-KB p65 (NF-KB p65)
Description: Quantitative sandwich ELISA for measuring Human Nuclear factor-KB p65 (NF-KB p65) in samples from cell culture supernatants, serum, whole blood, plasma and other biological fluids.
ELISA kit for Mouse Nuclear factor-KB p65 (NF-KB p65)
Description: A polyclonal antibody raised in Rabbit that recognizes and binds to Human NF-KB p100 (Ab-869). This antibody is tested and proven to work in the following applications:
Description: NF-kappaB is a pleiotropic transcription factor present in almost all cell types and is the endpoint of a series of signal transduction events that are initiated by a vast array of stimuli related to many biological processes such as inflammation, immunity, differentiation, cell growth, tumorigenesis and apoptosis. NF-kappaB is a homo- or heterodimeric complex formed by the Rel-like domain-containing proteins RELA/p65, RELB, NFKB1/p105, NFKB1/p50, REL and NFKB2/p52. The heterodimeric p65-p50 complex is the most abundant complex. The dimers bind at kappaB sites in the DNA of their target genes and the individual dimers have distinct preferences for different kappaB sites that they can bind with distinguishable affinity and specificity. Different dimer combinations act as transcriptional activators or repressors, respectively. NF-kappaB complexes are held in the cytoplasm in an inactive state complexed with members of the NF-kappaB inhibitor (I-kappaB) family. In a conventional activation pathway, I-kappaB is phosphorylated by I-kappaB kinases (IKKs) in response to different activators, subsequently degraded thus liberating the active NF-kappaB complex which translocates to the nucleus. NF-kappaB heterodimeric p65-p50 and p65-c-Rel complexes are transcriptional activators. The NF-kappaB p65-p65 complex appears to be involved in invasin-mediated activation of IL-8 expression. p65 shows a weak DNA-binding site which could contribute directly to DNA binding in the NF-kappaB complex.
Description: NF-kappaB is a pleiotropic transcription factor present in almost all cell types and is the endpoint of a series of signal transduction events that are initiated by a vast array of stimuli related to many biological processes such as inflammation, immunity, differentiation, cell growth, tumorigenesis and apoptosis. NF-kappaB is a homo- or heterodimeric complex formed by the Rel-like domain-containing proteins RELA/p65, RELB, NFKB1/p105, NFKB1/p50, REL and NFKB2/p52. The heterodimeric p65-p50 complex is the most abundant complex. The dimers bind at kappaB sites in the DNA of their target genes and the individual dimers have distinct preferences for different kappaB sites that they can bind with distinguishable affinity and specificity. Different dimer combinations act as transcriptional activators or repressors, respectively. NF-kappaB complexes are held in the cytoplasm in an inactive state complexed with members of the NF-kappaB inhibitor (I-kappaB) family. In a conventional activation pathway, I-kappaB is phosphorylated by I-kappaB kinases (IKKs) in response to different activators, subsequently degraded thus liberating the active NF-kappaB complex which translocates to the nucleus. NF-kappaB heterodimeric p65-p50 and RelB-p50 complexes are transcriptional activators. The NF-kappaB p50-p50 homodimer is a transcriptional repressor, but can act as a transcriptional activator when associated with BCL3.
Description: Recombinant Jurkat T cell expressing firefly luciferase gene under the control of NF-kB response elements with constitutive expression of human CD27 (also known as Tumor Necrosis Factor Receptor Superfamily Member 7, TNFRSF7, and T14, Genbank Accession #BC012160)
Description: A Mouse Monoclonal antibody against Acetyl NF kB P65?K314/K315) (5G11) from Human/ Mouse/ Rat. This antibody is tested and validated for IHC
Description: A Mouse Monoclonal antibody against Acetyl NF kB P65?K314/K315) (5G11) from Human/ Mouse/ Rat. This antibody is tested and validated for IHC
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Notably, ANKRD30B, a promising hub gene, was considerably hypermethylated in blood and downregulated in mind tissue from people with WS. Most CpG websites of ANKRD30B in blood have been considerably correlated with mind areas.
Furthermore, analyses of gene regulatory networks (GRNs) yielded grasp regulator transcription elements related to WS. Taken collectively, this systems-level method highlights the position of epigenetics in WS, and offers a doable rationalization for the advanced phenotypes noticed in sufferers with WS.
