The Experts below are selected from a list of 81156 Experts worldwide ranked by ideXlab platform
Naomi Kondo - One of the best experts on this subject based on the ideXlab platform.
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a novel mutation c 951c t in an exonic Splicing enhancer results in exon 10 skipping in the human mitochondrial acetoacetyl coa thiolase Gene
Molecular Genetics and Metabolism, 2010Co-Authors: Toshiyuki Fukao, Reiko Horikawa, Yasuhiro Naiki, Toju Tanaka, Masaki Takayanagi, Seiji Yamaguchi, Naomi KondoAbstract:Abstract Mitochondrial acetoacetyl-CoA thiolase (T2) deficiency is an inherited disorder affecting isoleucine catabolism and ketone body metabolism. A Japanese female developed a severe ketoacidotic attack at the age of 7 months. Urinary organic acid analysis showed elevated excretion of 2-methyl-3-hydroxybutyrate but not tiglylglycine. She was diagnosed as having T2 deficiency by enzyme assay using fibroblasts. Mutation analysis revealed a compound heterozygote of c.556G>T(D186Y) and c.951C>T( D317D ). Since c.951C>T does not cause amino acid change, we performed cDNA analysis and found that exon 10 skipping had occurred in the c.951C>T allele. A computer search using an ESE finder showed that an exonic Splicing enhancer sequence, SF2/ASF, was located in CTGA 951 C GC. We hypothesized that the exonic Splicing enhancer is necessary for accurate Splicing since the first nucleotide of exon 10 is C, which weakens the splice acceptor site of intron 9. We made a mini Gene construct including exon 9-truncated intron 9-exon 10-truncated intron 10-exon 11 for a Splicing experiment. We also made three mutant constructs which alter the SF2/ASF site (947C>T, 951C>T , 952G>A). An min-Gene Splicing experiment clearly showed that exon 10 skipping was induced in all three mutant constructs. Moreover, additional substitution of G for C at the first nucleotide of exon 10 resulted in normal Splicing in these three mutants. These results confirmed that c.951C>T diminished the effect of the exonic Splicing enhancer and caused exon 10 skipping.
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a novel single base substitution c 1124a g that activates a 5 base upstream cryptic splice donor site within exon 11 in the human mitochondrial acetoacetyl coa thiolase Gene
Molecular Genetics and Metabolism, 2008Co-Authors: Toshiyuki Fukao, Avihu Boneh, Yusuke Aoki, Naomi KondoAbstract:Abstract Most mutations related to aberrant Splicing occur in conserved splice acceptor and donor sites. Some exonic mutations also affect Splicing. We identified and characterized a point mutation (c.1124A>G) in an Australian patient (GK43) with mitochondrial acetoacetyl-CoA thiolase (T2) deficiency. GK43 is a homozygote of c.1124A>G, which activates a cryptic splice donor site 5 bases upstream from c.1124A>G within exon 11, causing aberrant Splicing in most transcripts. The aberrant Splicing results in c.1120–1163 (44-base) deletion, causing a frameshift in T2 mRNA. A mini-Gene Splicing experiment confirmed that the c.1124A>G substitution was responsible for this aberrant Splicing. This cryptic splice site has a Shapiro and Senapathy score (70.0) in a normal sequence but if mutated, the score (84.3) becomes higher than the one in the authentic splice donor site of intron 11 (81.4). This is an example in which a point mutation activates a cryptic splice donor site motif that is used preferentially over a downstream authentic splice site.
Toshiyuki Fukao - One of the best experts on this subject based on the ideXlab platform.
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a novel mutation c 951c t in an exonic Splicing enhancer results in exon 10 skipping in the human mitochondrial acetoacetyl coa thiolase Gene
Molecular Genetics and Metabolism, 2010Co-Authors: Toshiyuki Fukao, Reiko Horikawa, Yasuhiro Naiki, Toju Tanaka, Masaki Takayanagi, Seiji Yamaguchi, Naomi KondoAbstract:Abstract Mitochondrial acetoacetyl-CoA thiolase (T2) deficiency is an inherited disorder affecting isoleucine catabolism and ketone body metabolism. A Japanese female developed a severe ketoacidotic attack at the age of 7 months. Urinary organic acid analysis showed elevated excretion of 2-methyl-3-hydroxybutyrate but not tiglylglycine. She was diagnosed as having T2 deficiency by enzyme assay using fibroblasts. Mutation analysis revealed a compound heterozygote of c.556G>T(D186Y) and c.951C>T( D317D ). Since c.951C>T does not cause amino acid change, we performed cDNA analysis and found that exon 10 skipping had occurred in the c.951C>T allele. A computer search using an ESE finder showed that an exonic Splicing enhancer sequence, SF2/ASF, was located in CTGA 951 C GC. We hypothesized that the exonic Splicing enhancer is necessary for accurate Splicing since the first nucleotide of exon 10 is C, which weakens the splice acceptor site of intron 9. We made a mini Gene construct including exon 9-truncated intron 9-exon 10-truncated intron 10-exon 11 for a Splicing experiment. We also made three mutant constructs which alter the SF2/ASF site (947C>T, 951C>T , 952G>A). An min-Gene Splicing experiment clearly showed that exon 10 skipping was induced in all three mutant constructs. Moreover, additional substitution of G for C at the first nucleotide of exon 10 resulted in normal Splicing in these three mutants. These results confirmed that c.951C>T diminished the effect of the exonic Splicing enhancer and caused exon 10 skipping.
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a novel single base substitution c 1124a g that activates a 5 base upstream cryptic splice donor site within exon 11 in the human mitochondrial acetoacetyl coa thiolase Gene
Molecular Genetics and Metabolism, 2008Co-Authors: Toshiyuki Fukao, Avihu Boneh, Yusuke Aoki, Naomi KondoAbstract:Abstract Most mutations related to aberrant Splicing occur in conserved splice acceptor and donor sites. Some exonic mutations also affect Splicing. We identified and characterized a point mutation (c.1124A>G) in an Australian patient (GK43) with mitochondrial acetoacetyl-CoA thiolase (T2) deficiency. GK43 is a homozygote of c.1124A>G, which activates a cryptic splice donor site 5 bases upstream from c.1124A>G within exon 11, causing aberrant Splicing in most transcripts. The aberrant Splicing results in c.1120–1163 (44-base) deletion, causing a frameshift in T2 mRNA. A mini-Gene Splicing experiment confirmed that the c.1124A>G substitution was responsible for this aberrant Splicing. This cryptic splice site has a Shapiro and Senapathy score (70.0) in a normal sequence but if mutated, the score (84.3) becomes higher than the one in the authentic splice donor site of intron 11 (81.4). This is an example in which a point mutation activates a cryptic splice donor site motif that is used preferentially over a downstream authentic splice site.
Reisa A. Sperling - One of the best experts on this subject based on the ideXlab platform.
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Genetics of Gene Expression in the Aging Human Brain Reveal TDP-43 Proteinopathy Pathophysiology
Neuron, 2020Co-Authors: Hyun-sik Yang, Charles C. White, Hans-ulrich Klein, Chris Gaiteri, Daniel Felsky, Sara Mostafavi, Vladislav A. Petyuk, Reisa A. SperlingAbstract:Summary Here, we perform a genome-wide screen for variants that regulate the expression of Gene co-expression modules in the aging human brain; we discover and replicate such variants in the TMEM106B and RBFOX1 loci. The TMEM106B haplotype is known to influence the accumulation of TAR DNA-binding protein 43 kDa (TDP-43) proteinopathy, and the haplotype's large-scale transcriptomic effects include the dysregulation of lysosomal Genes and alterations in synaptic Gene Splicing that are also seen in the pathophysiology of TDP-43 proteinopathy. Further, a variant near GRN, another TDP-43 proteinopathy susceptibility Gene, shows concordant effects with the TMEM106B haplotype. Leveraging neuropathology data from the same participants, we also show that TMEM106B and APOE-amyloid-β effects converge to alter myelination and lysosomal Gene expression, which then contributes to TDP-43 accumulation. These results advance our mechanistic understanding of the TMEM106B TDP-43 risk haplotype and uncover a transcriptional program that mediates the converging effects of APOE-amyloid-β and TMEM106B on TDP-43 aggregation in older adults.
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Genetics of Gene Expression in the Aging Human Brain Reveal TDP-43 Proteinopathy Pathophysiology
SSRN Electronic Journal, 2019Co-Authors: Hyun-sik Yang, Charles C. White, Hans-ulrich Klein, Chris Gaiteri, Daniel Felsky, Sara Mostafavi, Vladislav A. Petyuk, Reisa A. SperlingAbstract:Genetic variants that control multiple distant Gene expressions are highly relevant for disease mechanisms. Here, in the aging human brain, we perform genome-wide screen of variants that regulate the average expression of each Gene co-expression network and identify such variants in TMEM106B and RBFOX1 loci. The TMEM106 Blocus coincides with a known TDP-43 proteinopathy risk haplotype, and shows large-scale transcriptomic effects including lysosomal dysregulation and altered synaptic Gene Splicing, that capture TDP-43 proteinopathy pathophysiology. Further, a locus in GRN, another TDP-43 proteinopathy risk Gene, shows concordant effects with the TMEM106B locus. Leveraging neuropathology data from the same participants, we also show that TMEM106B and APOE/amyloid-β effects converge to alter myelination/lysosomal Gene expression, which then contributes to TDP-43 accumulation. These results advance our mechanistic understanding of the TMEM106B TDP-43 risk allele, and uncover a transcriptional program that mediates APOE/amyloid-β and TMEM106B effects on TDP-43 aggregation in older adults.
Tao Jiang - One of the best experts on this subject based on the ideXlab platform.
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functional characterization of argininosuccinate lyase Gene variants by mini Gene Splicing assay
Frontiers in Genetics, 2019Co-Authors: Yanyun Wang, Yun Sun, Ming Liu, Xiaojuan Zhang, Tao JiangAbstract:Objective: To perform Gene detection and Gene mutation analysis in a family of inherited metabolic diseases with elevated CIT by MS/MS test. Methods: The peripheral blood samples were collected from the family members, and genomic DNA was extracted for Gene diagnosis using the total exon sequencing method. The novel mutation Gene was cloned into pEGFP-C1 vector, and the pathogenicity of the mutation was examined in cultured cells in vitro. Results: The clinical diagnosis of the proband as ASA was clear. Two pathogenic mutations, C.281G>T (p.Arg94Leu) and C.208-15 T>A were detected in the ASL Gene, and the two mutations had not been reported. The miniGene expression in vitro confirmed that C.208-15 T>A could cause aberrant Splicing, resulting in the retention of 13 bp in intron 2 to exon 3. In addition, both C.281G>T and C.208-15 T>A resulted in decreased EGFP expression in the vector pEGFP-C1. Conclusion: Two new pathogenic mutations of ASL Gene, C.208-15 T>A and C.281G>T, were found in an ASA family, which enriches the mutational profile of the ASL Gene and provides a basis for Genetic diagnosis of ASA. In addition to the mutation in the exon region, the base change in the intron may cause aberrant Splicing of mRNA and lead to abnormal structure and function of the protein.
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Image_1_Functional Characterization of Argininosuccinate Lyase Gene Variants by Mini-Gene Splicing Assay.TIF
2019Co-Authors: Yanyun Wang, Yun Sun, Ming Liu, Xiaojuan Zhang, Tao JiangAbstract:ObjectiveArgininosuccinate lyase (ASL) Gene mutations account for argininosuccinic aciduria (ASA). This study aimed to design a miniGene construct of ASL Gene in order to investigate the impact of variants on Splicing.MethodsThe peripheral blood samples were collected from the family members, and genomic DNA was extracted for Gene diagnosis using the total exon sequencing method. The novel mutation Gene was cloned into pEGFP-C1 vector, and the pathogenicity of the mutation was examined in cultured cells in vitro.ResultsThe clinical diagnosis of the proband as ASA was clear. Two pathogenic mutations, c.281G>T (p.Arg94Leu) and c.208-15 T>A were detected in the ASL Gene, and the two mutations had not been reported. The miniGene expression in vitro confirmed that c.208-15 T>A could cause aberrant Splicing, resulting in the retention of 13 bp in intron 2 to exon 3.ConclusionTwo new pathogenic mutations of ASL Gene, c.208-15 T>A and c.281G>T, were found in an ASA family, which enriches the mutational profile of the ASL Gene and provides a basis for Genetic diagnosis of ASA. MiniGenes are optimal approaches to determine whether the intron mutation can cause aberrant Splicing.
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Functional Characterization of Argininosuccinate Lyase Gene Variants by Mini-Gene Splicing Assay
Frontiers Media S.A., 2019Co-Authors: Yanyun Wang, Yun Sun, Ming Liu, Xiaojuan Zhang, Tao JiangAbstract:ObjectiveArgininosuccinate lyase (ASL) Gene mutations account for argininosuccinic aciduria (ASA). This study aimed to design a miniGene construct of ASL Gene in order to investigate the impact of variants on Splicing.MethodsThe peripheral blood samples were collected from the family members, and genomic DNA was extracted for Gene diagnosis using the total exon sequencing method. The novel mutation Gene was cloned into pEGFP-C1 vector, and the pathogenicity of the mutation was examined in cultured cells in vitro.ResultsThe clinical diagnosis of the proband as ASA was clear. Two pathogenic mutations, c.281G>T (p.Arg94Leu) and c.208-15 T>A were detected in the ASL Gene, and the two mutations had not been reported. The miniGene expression in vitro confirmed that c.208-15 T>A could cause aberrant Splicing, resulting in the retention of 13 bp in intron 2 to exon 3.ConclusionTwo new pathogenic mutations of ASL Gene, c.208-15 T>A and c.281G>T, were found in an ASA family, which enriches the mutational profile of the ASL Gene and provides a basis for Genetic diagnosis of ASA. MiniGenes are optimal approaches to determine whether the intron mutation can cause aberrant Splicing
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Table_1_Functional Characterization of Argininosuccinate Lyase Gene Variants by Mini-Gene Splicing Assay.docx
2019Co-Authors: Yanyun Wang, Yun Sun, Ming Liu, Xiaojuan Zhang, Tao JiangAbstract:ObjectiveArgininosuccinate lyase (ASL) Gene mutations account for argininosuccinic aciduria (ASA). This study aimed to design a miniGene construct of ASL Gene in order to investigate the impact of variants on Splicing.MethodsThe peripheral blood samples were collected from the family members, and genomic DNA was extracted for Gene diagnosis using the total exon sequencing method. The novel mutation Gene was cloned into pEGFP-C1 vector, and the pathogenicity of the mutation was examined in cultured cells in vitro.ResultsThe clinical diagnosis of the proband as ASA was clear. Two pathogenic mutations, c.281G>T (p.Arg94Leu) and c.208-15 T>A were detected in the ASL Gene, and the two mutations had not been reported. The miniGene expression in vitro confirmed that c.208-15 T>A could cause aberrant Splicing, resulting in the retention of 13 bp in intron 2 to exon 3.ConclusionTwo new pathogenic mutations of ASL Gene, c.208-15 T>A and c.281G>T, were found in an ASA family, which enriches the mutational profile of the ASL Gene and provides a basis for Genetic diagnosis of ASA. MiniGenes are optimal approaches to determine whether the intron mutation can cause aberrant Splicing.
Masutaka Furue - One of the best experts on this subject based on the ideXlab platform.
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an xpa Gene Splicing mutation resulting in trace protein expression in an elderly patient with xeroderma pigmentosum group a without neurological abnormalities
British Journal of Dermatology, 2017Co-Authors: Yoshito Takahashi, Y Endo, A Kusakakikushima, S Nakamaura, Yuka Nakazawa, Miki Uryu, Gaku Tsuji, Masutaka Furue, Shinichi MoriwakiAbstract:A certain relationship between XPA Gene mutations and the severity of symptoms has been observed in patients with xeroderma pigmentosum group A (XP-A). Patients with mutations within the DNA-binding domain usually exhibit severe symptoms, whereas Splicing mutations in the same domain sometimes cause very mild symptoms. This inconsistency can be explained by a small amount of functional XPA protein produced from normally spliced transcripts. We herein report the case of an adult Japanese XP-A patient with unusually mild symptoms. We identified a homozygous c.529G>A mutation in exon 4 of the XPA Gene, which resulted in aberrant Splicing with a 29-bp deletion in exon 4 causing a frameshift. Intact mRNA was observable, but a Western blot analysis failed to detect any normal XPA protein. We therefore evaluated the DNA repair capacity in normal cells in which the XPA expression was artificially diminished. The repair capacity was still present in cells with trace levels of the XPA protein. The repair capacity of the cells derived from our patient with mild symptoms was poor by comparison, but still significant compared to that of the cells derived from an XP-A patient with severe symptoms. These results provide strong evidence that a trace level of XPA protein can still exert a relatively strong repair capacity, resulting in only a mild phenotype. This article is protected by copyright. All rights reserved.
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an xpa Gene Splicing mutation resulting in trace protein expression in an elderly patient with xeroderma pigmentosum group a without neurological abnormalities
British Journal of Dermatology, 2017Co-Authors: Yoshito Takahashi, Y Endo, A Kusakakikushima, S Nakamaura, Yuka Nakazawa, Miki Uryu, Gaku Tsuji, Tomoo Ogi, Masutaka FurueAbstract:A certain relationship between XPA Gene mutations and the severity of symptoms has been observed in patients with xeroderma pigmentosum group A (XP-A). Patients with mutations within the DNA-binding domain usually exhibit severe symptoms, whereas Splicing mutations in the same domain sometimes cause very mild symptoms. This inconsistency can be explained by a small amount of functional XPA protein produced from normally spliced transcripts. We herein report the case of an adult Japanese patient with XP-A with unusually mild symptoms. We identified a homozygous c.529G>A mutation in exon 4 of the XPA Gene, which resulted in aberrant Splicing with a 29-bp deletion in exon 4 causing a frameshift. Intact mRNA was observable, but a Western blot analysis failed to detect any normal XPA protein. We therefore evaluated the DNA repair capacity in normal cells in which the XPA expression was artificially diminished. The repair capacity was still present in cells with trace levels of the XPA protein. The repair capacity of the cells derived from our patient with mild symptoms was poor by comparison, but still significant compared with that of the cells derived from a patient with XP-A with severe symptoms. These results provide strong evidence that a trace level of XPA protein can still exert a relatively strong repair capacity, resulting in only a mild phenotype.