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<t>Monoallelic</t> <t>methylation</t> of the human DNMT1 gene. A , location of the DNMT1 CpG island/promoter <t>DNA</t> methylation assays used for bisulfite sequencing. Genomic coordinates on chromosome 19 are shown, along with location of individual CpG dinucleotides ( dashes ) and CpG island location ( gray bar ) in relation to the DNMT 1 gene (exon 1 shown as a blue box , intron shown as an arrowed line ) according to the University of California Santa Cruz genome browser (hg_18 assembly). The arrows denote transcriptional direction. The location of rs8112895 SNP in the DNMT1_assay_4 is also shown. B , representative DNA methylation of a single full term human placental sample for assay_1 ( i ) and assay_4 ( ii ) showing a monoallelic pattern of methylation within the human placenta. Between eight and 12 individual clones were sequenced for each sample. The circles correspond to CpG sites denoted by black dashes in A. Closed circles , methylation; open circles , lack of methylation. Mean methylation levels seen at each CpG site in multiple placental samples for each assay are shown as shaded bars between 0 and 100% ( n = 4). Missing circles indicate CpG sites for which no information was obtained. C , DNMT1_4 methylation assay spanning SNP rs8112895 was used for bisulfite sequencing methylation analysis in purified first trimester cytotrophoblast cells. i , reference genomic DNA sequence surrounding rs8112895 SNP ( R ) and CpG dinucleotides ( boxed ). ii , individual clone sequences showing the presence of bisulfite-converted CG sites ( TG ) and unconverted CG sites ( CG ) in both maternal and paternal alleles from this heterozygous sample. This confirms a lack of parent of origin of the observed DNMT1 promoter methylation.
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Summary of studies (from the past 10 years) on the effect of pre-gestational and prenatal cannabis exposure on genes and molecular pathways in humans. Studies are in reverse order chronologically by publication year

Journal: Current Environmental Health Reports

Article Title: Cannabis Exposure During Critical Windows of Development: Epigenetic and Molecular Pathways Implicated in Neuropsychiatric Disease

doi: 10.1007/s40572-020-00275-4

Figure Lengend Snippet: Summary of studies (from the past 10 years) on the effect of pre-gestational and prenatal cannabis exposure on genes and molecular pathways in humans. Studies are in reverse order chronologically by publication year

Article Snippet: Fransquet et al. 2017 [ •] , Nested case-control study n = 804 maternal subjects; 44 cannabis users anytime during pregnancy and 760 non-user controls , Prenatal , Buccal cells of neonates of maternal subjects F1 generation , DNA methylation SEQUENOM MassARRAY , • Gestational cannabis use associated with increased methylation at one CpG site tested in DRD4 (CpG.21.22.2) (β + 1.48, 95% CI: 0.02–2.93, p = 0.047). • At CpG.32, weak evidence that gestational cannabis is associated with increased methylation when adjusting for other substance use (β + .67, 95% CI: − 0.12-1.46, p = 0.09). • No associations remained significant after correction for multiple testing using a Bonferroni corrected significance level of 0.0026 given the 19 CpG units examined. , • DRD4 plays key role in dopamine signaling and is associated with drug use. • Increased DRD4 methylation in saliva has been associated with increased severity of ADHD symptoms in children. • Tissue specificity (using buccal cells instead of brain tissue) could have contributed to the null findings..

Techniques: Cannabis, DNA Methylation Assay, Expressing, Methylation, Immunoprecipitation, Methylated DNA Immunoprecipitation, Control, Methylation Sequencing, Membrane, Gene Expression

Summary of studies (from the past 10 years) on the effect of pre-gestational, prenatal, and adolescent cannabis exposure on genes and molecular pathways in animals. Studies are in reverse order chronologically by publication year

Journal: Current Environmental Health Reports

Article Title: Cannabis Exposure During Critical Windows of Development: Epigenetic and Molecular Pathways Implicated in Neuropsychiatric Disease

doi: 10.1007/s40572-020-00275-4

Figure Lengend Snippet: Summary of studies (from the past 10 years) on the effect of pre-gestational, prenatal, and adolescent cannabis exposure on genes and molecular pathways in animals. Studies are in reverse order chronologically by publication year

Article Snippet: Fransquet et al. 2017 [ •] , Nested case-control study n = 804 maternal subjects; 44 cannabis users anytime during pregnancy and 760 non-user controls , Prenatal , Buccal cells of neonates of maternal subjects F1 generation , DNA methylation SEQUENOM MassARRAY , • Gestational cannabis use associated with increased methylation at one CpG site tested in DRD4 (CpG.21.22.2) (β + 1.48, 95% CI: 0.02–2.93, p = 0.047). • At CpG.32, weak evidence that gestational cannabis is associated with increased methylation when adjusting for other substance use (β + .67, 95% CI: − 0.12-1.46, p = 0.09). • No associations remained significant after correction for multiple testing using a Bonferroni corrected significance level of 0.0026 given the 19 CpG units examined. , • DRD4 plays key role in dopamine signaling and is associated with drug use. • Increased DRD4 methylation in saliva has been associated with increased severity of ADHD symptoms in children. • Tissue specificity (using buccal cells instead of brain tissue) could have contributed to the null findings..

Techniques: Cannabis, Animal Model, DNA Methylation Assay, Methylated DNA Immunoprecipitation, Immunoprecipitation, Methylation, Expressing, Gene Expression, Mouse Assay, Enzyme-linked Immunosorbent Assay, Control, Modification, Functional Assay, Disruption, Activity Assay, Methylation Sequencing, Virus, Quantitative Proteomics, Cell Function Assay, Comparison, Membrane, Transmission Assay, Binding Assay, Inhibition, Chromatin Immunoprecipitation

Summary of main projects in the PANINI network

Journal: Aging Clinical and Experimental Research

Article Title: Physical Activity and Nutrition INfluences In ageing (PANINI): consortium mission statement

doi: 10.1007/s40520-017-0823-7

Figure Lengend Snippet: Summary of main projects in the PANINI network

Article Snippet: 10. University of Bologna , Epigenetics of nutrition in ageing , DNA methylation analysis (gene-targeted) by Sequenom ® MassARRAY EpiTYPER platform , NU-AGE samples and new PANINI data , Changes in DNA methylation patterns induced by dietary and physical interventions. Development of a gene-targeted epigenetic clock.

Techniques: Battery, Activity Assay, Modification, DNA Methylation Assay, Variant Assay

Monoallelic methylation of the human DNMT1 gene. A , location of the DNMT1 CpG island/promoter DNA methylation assays used for bisulfite sequencing. Genomic coordinates on chromosome 19 are shown, along with location of individual CpG dinucleotides ( dashes ) and CpG island location ( gray bar ) in relation to the DNMT 1 gene (exon 1 shown as a blue box , intron shown as an arrowed line ) according to the University of California Santa Cruz genome browser (hg_18 assembly). The arrows denote transcriptional direction. The location of rs8112895 SNP in the DNMT1_assay_4 is also shown. B , representative DNA methylation of a single full term human placental sample for assay_1 ( i ) and assay_4 ( ii ) showing a monoallelic pattern of methylation within the human placenta. Between eight and 12 individual clones were sequenced for each sample. The circles correspond to CpG sites denoted by black dashes in A. Closed circles , methylation; open circles , lack of methylation. Mean methylation levels seen at each CpG site in multiple placental samples for each assay are shown as shaded bars between 0 and 100% ( n = 4). Missing circles indicate CpG sites for which no information was obtained. C , DNMT1_4 methylation assay spanning SNP rs8112895 was used for bisulfite sequencing methylation analysis in purified first trimester cytotrophoblast cells. i , reference genomic DNA sequence surrounding rs8112895 SNP ( R ) and CpG dinucleotides ( boxed ). ii , individual clone sequences showing the presence of bisulfite-converted CG sites ( TG ) and unconverted CG sites ( CG ) in both maternal and paternal alleles from this heterozygous sample. This confirms a lack of parent of origin of the observed DNMT1 promoter methylation.

Journal: The Journal of Biological Chemistry

Article Title: DNA Methylation-mediated Down-regulation of DNA Methyltransferase-1 ( DNMT1 ) Is Coincident with, but Not Essential for, Global Hypomethylation in Human Placenta

doi: 10.1074/jbc.M109.064956

Figure Lengend Snippet: Monoallelic methylation of the human DNMT1 gene. A , location of the DNMT1 CpG island/promoter DNA methylation assays used for bisulfite sequencing. Genomic coordinates on chromosome 19 are shown, along with location of individual CpG dinucleotides ( dashes ) and CpG island location ( gray bar ) in relation to the DNMT 1 gene (exon 1 shown as a blue box , intron shown as an arrowed line ) according to the University of California Santa Cruz genome browser (hg_18 assembly). The arrows denote transcriptional direction. The location of rs8112895 SNP in the DNMT1_assay_4 is also shown. B , representative DNA methylation of a single full term human placental sample for assay_1 ( i ) and assay_4 ( ii ) showing a monoallelic pattern of methylation within the human placenta. Between eight and 12 individual clones were sequenced for each sample. The circles correspond to CpG sites denoted by black dashes in A. Closed circles , methylation; open circles , lack of methylation. Mean methylation levels seen at each CpG site in multiple placental samples for each assay are shown as shaded bars between 0 and 100% ( n = 4). Missing circles indicate CpG sites for which no information was obtained. C , DNMT1_4 methylation assay spanning SNP rs8112895 was used for bisulfite sequencing methylation analysis in purified first trimester cytotrophoblast cells. i , reference genomic DNA sequence surrounding rs8112895 SNP ( R ) and CpG dinucleotides ( boxed ). ii , individual clone sequences showing the presence of bisulfite-converted CG sites ( TG ) and unconverted CG sites ( CG ) in both maternal and paternal alleles from this heterozygous sample. This confirms a lack of parent of origin of the observed DNMT1 promoter methylation.

Article Snippet: Mass spectrometry-based DNA methylation profiling (SEQUENOM MassARRAY EPITYPER) revealed an absence of methylation of DNMT3A or -3B genes in all human tissues, including placenta and purified cytotrophoblasts ( supplemental Fig. 1 ).

Techniques: Methylation, DNA Methylation Assay, Methylation Sequencing, Clone Assay, Purification, Sequencing

DNMT1 promoter methylation is restricted to primates. Bisulfite DNA sequencing confirmed promoter methylation of the DNMT1 gene in full term baboon ( A ) and marmoset ( B ) placental tissue, with no evidence for methylation in mouse ( C ), bovine, or guinea pig placental tissue (data not shown).

Journal: The Journal of Biological Chemistry

Article Title: DNA Methylation-mediated Down-regulation of DNA Methyltransferase-1 ( DNMT1 ) Is Coincident with, but Not Essential for, Global Hypomethylation in Human Placenta

doi: 10.1074/jbc.M109.064956

Figure Lengend Snippet: DNMT1 promoter methylation is restricted to primates. Bisulfite DNA sequencing confirmed promoter methylation of the DNMT1 gene in full term baboon ( A ) and marmoset ( B ) placental tissue, with no evidence for methylation in mouse ( C ), bovine, or guinea pig placental tissue (data not shown).

Article Snippet: Mass spectrometry-based DNA methylation profiling (SEQUENOM MassARRAY EPITYPER) revealed an absence of methylation of DNMT3A or -3B genes in all human tissues, including placenta and purified cytotrophoblasts ( supplemental Fig. 1 ).

Techniques: Methylation, DNA Sequencing