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[PMID]:29179671
[Au] Autor:Hudaiberdiev S; Shmakov S; Wolf YI; Terns MP; Makarova KS; Koonin EV
[Ad] Endereço:National Center for Biotechnology Information, National Institutes of Health, Bethesda, MD, USA.
[Ti] Título:Phylogenomics of Cas4 family nucleases.
[So] Source:BMC Evol Biol;17(1):232, 2017 Nov 28.
[Is] ISSN:1471-2148
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:BACKGROUND: The Cas4 family endonuclease is a component of the adaptation module in many variants of CRISPR-Cas adaptive immunity systems. Unlike most of the other Cas proteins, Cas4 is often encoded outside CRISPR-cas loci (solo-Cas4) and is also found in mobile genetic elements (MGE-Cas4). RESULTS: As part of our ongoing investigation of CRISPR-Cas evolution, we explored the phylogenomics of the Cas4 family. About 90% of the archaeal genomes encode Cas4 compared to only about 20% of the bacterial genomes. Many archaea encode both the CRISPR-associated form (CAS-Cas4) and solo-Cas4, whereas in bacteria, this combination is extremely rare. The solo-cas4 genes are over-represented in environmental bacteria and archaea with small genomes that typically lack CRISPR-Cas, suggesting that Cas4 could perform uncharacterized defense or repair functions in these microbes. Phylogenomic analysis indicates that both the CRISPR-associated cas4 genes are often transferred horizontally but almost exclusively, as part of the adaptation module. The evolutionary integrity of the adaptation module sharply contrasts the rampant shuffling of CRISPR-cas modules whereby a given variant of the adaptation module can combine with virtually any effector module. The solo-cas4 genes evolve primarily via vertical inheritance and are subject only to occasional horizontal transfer. The selection pressure on cas4 genes does not substantially differ between CAS-Cas4 and solo-cas4, and is close to the genomic median. Thus, cas4 genes, similarly to cas1 and cas2, evolve similarly to 'regular' microbial genes involved in various cellular functions, showing no evidence of direct involvement in virus-host arms races. A notable feature of the Cas4 family evolution is the frequent recruitment of cas4 genes by various mobile genetic elements (MGE), particularly, archaeal viruses. The functions of Cas4 in these elements are unknown and potentially might involve anti-defense roles. CONCLUSIONS: Unlike most of the other Cas proteins, Cas4 family members are as often encoded by stand-alone genes as they are incorporated in CRISPR-Cas systems. In addition, cas4 genes were repeatedly recruited by MGE, perhaps, for anti-defense functions. Experimental characterization of the solo and MGE-encoded Cas4 nucleases is expected to reveal currently uncharacterized defense and anti-defense systems and their interactions with CRISPR-Cas systems.
[Mh] Termos MeSH primário: Sistemas CRISPR-Cas/genética
Endonucleases/genética
Genômica
Família Multigênica
[Mh] Termos MeSH secundário: Archaea/enzimologia
Archaea/genética
Bactérias/enzimologia
Bactérias/genética
Sequência de Bases
Elementos de DNA Transponíveis/genética
Transferência Genética Horizontal/genética
Loci Gênicos
Genoma Arqueal
Genoma Bacteriano
Filogenia
Seleção Genética
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (DNA Transposable Elements); EC 3.1.- (Endonucleases)
[Em] Mês de entrada:1803
[Cu] Atualização por classe:180309
[Lr] Data última revisão:
180309
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171129
[St] Status:MEDLINE
[do] DOI:10.1186/s12862-017-1081-1


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[PMID]:29205966
[Au] Autor:Deng XD; Zhang W; Zhang B; Ma Y; Muer CE; Zhang LX; Xie Y; Liu Y
[Ad] Endereço:Department of Forensic Medicine, North Sichuan Medical College, Nanchong 637000, China.
[Ti] Título:[Expression of XPG Gene in Forensic Age Estimation].
[So] Source:Fa Yi Xue Za Zhi;32(6):415-419, 2016 Dec.
[Is] ISSN:1004-5619
[Cp] País de publicação:China
[La] Idioma:chi
[Ab] Resumo:OBJECTIVES: To explore the expression of xeroderma pigmentosum complementation group G ( ) gene in healthy Han population of different ages and to analysis the relationship between the mRNA and protein expression levels of XPG and age, which may provide a new molecular-biological indicator for forensic age determination. METHODS: Total 150 samples of peripheral blood were collected from healthy Han population of different ages. Total RNA of peripheral blood mononuclear cell (PBMC) were extracted by TRIzol method, and the relative expression of mRNA in PBMC was detected by quantitative real-time PCR, and the protein expression levels of XPG in plasma were detected by ELISA. RESULTS: The mRNA and protein expression levels of XPG in ≤18 years old group were significantly different from 19-45 years old group and ≥46 years old group ( <0.05), while there was no significant difference between 19-45 years old group and ≥46 years old group ( >0.05). No significant sex differences were observed in mRNA and protein expression levels of XPG ( >0.05). CONCLUSIONS: The relative expression level of mRNA in PBMC declines with the increase of age in younger age, while the protein expression level in plasma increases with age, and gene can be used as one of new markers for forensic age estimation.
[Mh] Termos MeSH primário: Fatores Etários
Proteínas de Ligação a DNA/genética
Endonucleases/genética
Proteínas Nucleares/genética
Fatores de Transcrição/genética
[Mh] Termos MeSH secundário: Adulto
Grupo com Ancestrais do Continente Asiático
Genética Forense
Seres Humanos
Leucócitos Mononucleares
Meia-Idade
RNA Mensageiro
Reação em Cadeia da Polimerase em Tempo Real
Adulto Jovem
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (DNA excision repair protein ERCC-5); 0 (DNA-Binding Proteins); 0 (Nuclear Proteins); 0 (RNA, Messenger); 0 (Transcription Factors); EC 3.1.- (Endonucleases)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180308
[Lr] Data última revisão:
180308
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171206
[St] Status:MEDLINE
[do] DOI:10.3969/j.issn.1004-5619.2016.06.005


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[PMID]:29379011
[Au] Autor:Yoon Y; Wang D; Tai PWL; Riley J; Gao G; Rivera-Pérez JA
[Ad] Endereço:Department of Pediatrics, Division of Genes and Development, University of Massachusetts Medical School, 55 Lake Avenue North, Worcester, MA, 01655, USA.
[Ti] Título:Streamlined ex vivo and in vivo genome editing in mouse embryos using recombinant adeno-associated viruses.
[So] Source:Nat Commun;9(1):412, 2018 01 29.
[Is] ISSN:2041-1723
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Recent advances using CRISPR-Cas9 approaches have dramatically enhanced the ease for genetic manipulation in rodents. Notwithstanding, the methods to deliver nucleic acids into pre-implantation embryos have hardly changed since the original description of mouse transgenesis more than 30 years ago. Here we report a novel strategy to generate genetically modified mice by transduction of CRISPR-Cas9 components into pre-implantation mouse embryos via recombinant adeno-associated viruses (rAAVs). Using this approach, we efficiently generated a variety of targeted mutations in explanted embryos, including indel events produced by non-homologous end joining and tailored mutations using homology-directed repair. We also achieved gene modification in vivo by direct delivery of rAAV particles into the oviduct of pregnant females. Our approach greatly simplifies the generation of genetically modified mice and, more importantly, opens the door for streamlined gene editing in other mammalian species.
[Mh] Termos MeSH primário: Sistemas CRISPR-Cas
Repetições Palindrômicas Curtas Agrupadas e Regularmente Espaçadas
Dependovirus/genética
Desenvolvimento Embrionário/genética
Edição de Genes/métodos
Engenharia Genética/métodos
[Mh] Termos MeSH secundário: Animais
Proteínas de Bactérias/genética
Proteínas de Bactérias/metabolismo
Blastocisto
Reparo do DNA por Junção de Extremidades
Dependovirus/metabolismo
Endonucleases/genética
Endonucleases/metabolismo
Tubas Uterinas/embriologia
Feminino
Genes Reporter
Proteínas de Fluorescência Verde/genética
Proteínas de Fluorescência Verde/metabolismo
Células HEK293
Seres Humanos
Camundongos
Camundongos Transgênicos
Mutagênese Sítio-Dirigida
Gravidez
Reparo de DNA por Recombinação
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, N.I.H., EXTRAMURAL
[Nm] Nome de substância:
0 (Bacterial Proteins); 0 (enhanced green fluorescent protein); 147336-22-9 (Green Fluorescent Proteins); EC 3.1.- (Cas9 endonuclease Streptococcus pyogenes); EC 3.1.- (Endonucleases)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180227
[Lr] Data última revisão:
180227
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:180131
[St] Status:MEDLINE
[do] DOI:10.1038/s41467-017-02706-7


  4 / 7878 MEDLINE  
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[PMID]:29390553
[Au] Autor:Ryu H; Song IC; Choi YS; Yun HJ; Jo DY; Kim JM; Ko YB; Lee HJ
[Ad] Endereço:Department of Internal Medicine.
[Ti] Título:ERCC1 expression status predicts the response and survival of patients with metastatic or recurrent cervical cancer treated via platinum-based chemotherapy.
[So] Source:Medicine (Baltimore);96(51):e9402, 2017 Dec.
[Is] ISSN:1536-5964
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:The deoxyribonucleic acid (DNA) repair gene encoding the excision-repair cross-complementation group 1 (ERCC1) protein is known to predict the response to platinum-based chemotherapy. Our aim was to explore whether ERCC1 expression predicted tumor response and survival in patients with recurrent or metastatic cervical cancer treated via platinum-based chemotherapy. We analyzed 32 such patients. ERCC1 expression was assessed immunohistochemically in pretreatment biopsy samples. Of the 32 patients, 13 (40.6%) were ERCC1 high. ERCC1-low patients exhibited a significantly higher response rate (73.7%) than did others (15.4%). The median progression-free survival differed significantly by ERCC1 status, being 135 days in ERCC1-high and 242 days in ERCC1-low patients (hazard ratio, 2.428; 95% confidence interval, 1.145-5.148, P = .032). Overall survival was significantly longer in ERCC1-low (617 days) than in ERCC1-high (320 days) patients (hazard ratio, 2.322; 95% confidence interval, 1.051-5.29; P = .037). Thus, pretreatment ERCC1 expression status can be used to predict tumor response and survival of patients with recurrent or metastatic uterine cervical cancer receiving platinum-based chemotherapy.
[Mh] Termos MeSH primário: Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico
Biomarcadores Tumorais/metabolismo
Carcinoma/tratamento farmacológico
Proteínas de Ligação a DNA/metabolismo
Endonucleases/metabolismo
Neoplasias do Colo do Útero/tratamento farmacológico
[Mh] Termos MeSH secundário: Adulto
Idoso
Idoso de 80 Anos ou mais
Antineoplásicos/uso terapêutico
Carboplatina/uso terapêutico
Carcinoma/metabolismo
Carcinoma/mortalidade
Carcinoma/secundário
Cisplatino/uso terapêutico
Feminino
Fluoruracila/uso terapêutico
Seguimentos
Seres Humanos
Imuno-Histoquímica
Meia-Idade
Recidiva Local de Neoplasia/tratamento farmacológico
Recidiva Local de Neoplasia/metabolismo
Recidiva Local de Neoplasia/mortalidade
Paclitaxel/uso terapêutico
Neoplasias Pélvicas/patologia
Estudos Retrospectivos
Análise de Sobrevida
Topotecan/uso terapêutico
Resultado do Tratamento
Neoplasias do Colo do Útero/metabolismo
Neoplasias do Colo do Útero/mortalidade
Neoplasias do Colo do Útero/secundário
[Pt] Tipo de publicação:JOURNAL ARTICLE; OBSERVATIONAL STUDY
[Nm] Nome de substância:
0 (Antineoplastic Agents); 0 (Biomarkers, Tumor); 0 (DNA-Binding Proteins); 7M7YKX2N15 (Topotecan); BG3F62OND5 (Carboplatin); EC 3.1.- (ERCC1 protein, human); EC 3.1.- (Endonucleases); P88XT4IS4D (Paclitaxel); Q20Q21Q62J (Cisplatin); U3P01618RT (Fluorouracil)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180214
[Lr] Data última revisão:
180214
[Sb] Subgrupo de revista:AIM; IM
[Da] Data de entrada para processamento:180203
[St] Status:MEDLINE
[do] DOI:10.1097/MD.0000000000009402


  5 / 7878 MEDLINE  
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[PMID]:29247747
[Au] Autor:Zhao M; Li S; Zhou L; Shen Q; Zhu H; Zhu X
[Ad] Endereço:Department of Obstetrics and Gynecology, the Second Affiliated Hospital of Wenzhou Medical University, Wenzhou, China.
[Ti] Título:Prognostic values of excision repair cross-complementing genes mRNA expression in ovarian cancer patients.
[So] Source:Life Sci;194:34-39, 2018 Feb 01.
[Is] ISSN:1879-0631
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:Excision repair cross-complementing (ERCC) genes, key components of the nucleotide excision repair pathway, are regarded as crucial factors for DNA repair capacity. Previous studies have investigated prognostic values of ERCC genes in a number of malignancies. However, the relationship between ERCC genes and prognosis of ovarian cancer patients remains controversial. Therefore, in the current study, we systematically analyze the prognostic values of ERCC genes in ovarian cancer by the Kaplan-Meier plotter, which includes updated gene expression data and survival information of 1656 ovarian cancer patients. Our results showed that high expression of ERCC1 and ERCC8 mRNA was related to a worse overall survival among ovarian cancer patients, especially in late stage and poor differentiation serous ovarian patients. Increased ERCC4 mRNA expression indicated a better overall survival among serous ovarian cancer patients. The other ERCC genes were uncorrelated with prognosis in ovarian cancer. These results indicate that some ERCC genes have critical prognostic values in ovarian cancer.
[Mh] Termos MeSH primário: Enzimas Reparadoras do DNA/genética
Reparo do DNA
Proteínas de Ligação a DNA/genética
Endonucleases/genética
Neoplasias Ovarianas/genética
RNA Mensageiro/genética
Fatores de Transcrição/genética
[Mh] Termos MeSH secundário: Feminino
Regulação Neoplásica da Expressão Gênica
Seres Humanos
Estimativa de Kaplan-Meier
Neoplasias Ovarianas/diagnóstico
Neoplasias Ovarianas/patologia
Ovário/metabolismo
Ovário/patologia
Prognóstico
Análise de Sobrevida
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (DNA-Binding Proteins); 0 (ERCC8 protein, human); 0 (RNA, Messenger); 0 (Transcription Factors); 0 (xeroderma pigmentosum group F protein); EC 3.1.- (ERCC1 protein, human); EC 3.1.- (Endonucleases); EC 6.5.1.- (DNA Repair Enzymes)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180213
[Lr] Data última revisão:
180213
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171217
[St] Status:MEDLINE


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[PMID]:28456968
[Au] Autor:Wang W; Zhang Y; Wang H
[Ad] Endereço:The Jackson Laboratory, Bar Harbor, ME, 04609, USA.
[Ti] Título:Generating Mouse Models Using Zygote Electroporation of Nucleases (ZEN) Technology with High Efficiency and Throughput.
[So] Source:Methods Mol Biol;1605:219-230, 2017.
[Is] ISSN:1940-6029
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Mouse models with genetic modifications are widely used in biology and biomedical research. Although the application of CRISPR-Cas9 system greatly accelerated the process of generating genetically modified mice, the delivery method depending on manual injection of the components into the embryos remains a bottleneck, as it is laborious, low throughput, and technically demanding. To overcome this limitation, we invented and optimized the ZEN (Zygote electroporation of nucleases) technology to deliver CRISPR-Cas9 reagents via electroporation. Using ZEN, we were able to generate genetically modified mouse models with high efficiency and throughput. Here, we describe the protocol in great detail.
[Mh] Termos MeSH primário: Sistemas CRISPR-Cas
Eletroporação/métodos
Modelos Animais
[Mh] Termos MeSH secundário: Animais
Endonucleases/administração & dosagem
Camundongos
Camundongos Transgênicos
Zigoto
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
EC 3.1.- (Endonucleases)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180212
[Lr] Data última revisão:
180212
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170501
[St] Status:MEDLINE
[do] DOI:10.1007/978-1-4939-6988-3_15


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[PMID]:28462777
[Au] Autor:Lee K; Mackley VA; Rao A; Chong AT; Dewitt MA; Corn JE; Murthy N
[Ad] Endereço:GenEdit Inc, Berkeley, United States.
[Ti] Título:Synthetically modified guide RNA and donor DNA are a versatile platform for CRISPR-Cas9 engineering.
[So] Source:Elife;6, 2017 05 02.
[Is] ISSN:2050-084X
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Chemical modification of the gRNA and donor DNA has great potential for improving the gene editing efficiency of Cas9 and Cpf1, but has not been investigated extensively. In this report, we demonstrate that the gRNAs of Cas9 and Cpf1, and donor DNA can be chemically modified at their terminal positions without losing activity. Moreover, we show that 5' fluorescently labeled donor DNA can be used as a marker to enrich HDR edited cells by a factor of two through cell sorting. In addition, we demonstrate that the gRNA and donor DNA can be directly conjugated together into one molecule, and show that this gRNA-donor DNA conjugate is three times better at transfecting cells and inducing HDR, with cationic polymers, than unconjugated gRNA and donor DNA. The tolerance of the gRNA and donor DNA to chemical modifications has the potential to enable new strategies for genome engineering.
[Mh] Termos MeSH primário: Sistemas CRISPR-Cas
DNA/química
DNA/genética
Edição de Genes/métodos
RNA Guia/química
RNA Guia/genética
[Mh] Termos MeSH secundário: Proteínas de Bactérias/metabolismo
Endonucleases/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Bacterial Proteins); 0 (RNA, Guide); 9007-49-2 (DNA); EC 3.1.- (Cas9 endonuclease Streptococcus pyogenes); EC 3.1.- (Cpf1 nuclease, Acidaminococcus); EC 3.1.- (Endonucleases)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180211
[Lr] Data última revisão:
180211
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170503
[St] Status:MEDLINE


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[PMID]:29321179
[Au] Autor:Reginato G; Cannavo E; Cejka P
[Ad] Endereço:Institute for Research in Biomedicine, Università della Svizzera italiana, Bellinzona 6500, Switzerland.
[Ti] Título:Physiological protein blocks direct the Mre11-Rad50-Xrs2 and Sae2 nuclease complex to initiate DNA end resection.
[So] Source:Genes Dev;31(23-24):2325-2330, 2017 12 01.
[Is] ISSN:1549-5477
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:DNA double-strand break repair by homologous recombination is initiated by DNA end resection, which is commenced by the Mre11-Rad50-Xrs2 complex and Sae2 in yeast. Here we report that the nonhomologous end joining factor Ku limits the exonuclease activity of Mre11 and promotes its endonuclease to cleave 5'-terminated DNA strands at break sites. Following initial endonucleolytic cleavage past the obstacle, Exo1 specifically extends the resection track, leading to the generation of long 3' overhangs that are required for homologous recombination. These experiments provide mechanistic insights into how short-range and long-range DNA end resection enzymes overcome obstacles near broken DNA ends to initiate recombination.
[Mh] Termos MeSH primário: Reparo do DNA por Junção de Extremidades
Endonucleases/metabolismo
Exonucleases/metabolismo
Proteínas de Saccharomyces cerevisiae/metabolismo
Saccharomyces cerevisiae/fisiologia
[Mh] Termos MeSH secundário: Animais
Clivagem do DNA
Proteínas de Ligação a DNA/metabolismo
Endodesoxirribonucleases/metabolismo
Ativação Enzimática/genética
Exodesoxirribonucleases/metabolismo
Complexos Multiproteicos/metabolismo
Saccharomyces cerevisiae/enzimologia
Saccharomyces cerevisiae/genética
Proteínas de Saccharomyces cerevisiae/genética
Células Sf9
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (DNA-Binding Proteins); 0 (Multiprotein Complexes); 0 (RAD50 protein, S cerevisiae); 0 (SAE2 protein, S cerevisiae); 0 (Saccharomyces cerevisiae Proteins); 0 (XRS2 protein, S cerevisiae); 0 (high affinity DNA-binding factor, S cerevisiae); EC 3.1.- (Endodeoxyribonucleases); EC 3.1.- (Endonucleases); EC 3.1.- (Exodeoxyribonucleases); EC 3.1.- (Exonucleases); EC 3.1.- (MRE11 protein, S cerevisiae); EC 3.1.11.1 (exodeoxyribonuclease I)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180208
[Lr] Data última revisão:
180208
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:180112
[St] Status:MEDLINE
[do] DOI:10.1101/gad.308254.117


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[PMID]:29321177
[Au] Autor:Wang W; Daley JM; Kwon Y; Krasner DS; Sung P
[Ad] Endereço:Department of Molecular Biophysics and Biochemistry, Yale University School of Medicine, New Haven, Connecticut 06520, USA.
[Ti] Título:Plasticity of the Mre11-Rad50-Xrs2-Sae2 nuclease ensemble in the processing of DNA-bound obstacles.
[So] Source:Genes Dev;31(23-24):2331-2336, 2017 12 01.
[Is] ISSN:1549-5477
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:The budding yeast Mre11-Rad50-Xrs2 (MRX) complex and Sae2 function together in DNA end resection during homologous recombination. Here we show that the Ku complex shields DNA ends from exonucleolytic digestion but facilitates endonucleolytic scission by MRX with a dependence on ATP and Sae2. The incision site is enlarged into a DNA gap via the exonuclease activity of MRX, which is stimulated by Sae2 without ATP being present. RPA renders a partially resected or palindromic DNA structure susceptible to MRX-Sae2, and internal protein blocks also trigger DNA cleavage. We present models for how MRX-Sae2 creates entry sites for the long-range resection machinery.
[Mh] Termos MeSH primário: Reparo do DNA por Junção de Extremidades
Reparo do DNA/fisiologia
Endonucleases/metabolismo
Exonucleases/metabolismo
Complexos Multienzimáticos/metabolismo
Proteínas de Saccharomyces cerevisiae/metabolismo
Saccharomyces cerevisiae/fisiologia
[Mh] Termos MeSH secundário: Clivagem do DNA
Proteínas de Ligação a DNA/metabolismo
Endodesoxirribonucleases/metabolismo
Ativação Enzimática/genética
Exodesoxirribonucleases/metabolismo
Complexos Multiproteicos/metabolismo
Saccharomyces cerevisiae/enzimologia
Saccharomyces cerevisiae/genética
Proteínas de Saccharomyces cerevisiae/genética
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, N.I.H., EXTRAMURAL
[Nm] Nome de substância:
0 (DNA-Binding Proteins); 0 (Multienzyme Complexes); 0 (Multiprotein Complexes); 0 (RAD50 protein, S cerevisiae); 0 (SAE2 protein, S cerevisiae); 0 (Saccharomyces cerevisiae Proteins); 0 (XRS2 protein, S cerevisiae); 0 (high affinity DNA-binding factor, S cerevisiae); EC 3.1.- (Endodeoxyribonucleases); EC 3.1.- (Endonucleases); EC 3.1.- (Exodeoxyribonucleases); EC 3.1.- (Exonucleases); EC 3.1.- (MRE11 protein, S cerevisiae); EC 3.1.11.1 (exodeoxyribonuclease I)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180208
[Lr] Data última revisão:
180208
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:180112
[St] Status:MEDLINE
[do] DOI:10.1101/gad.307900.117


  10 / 7878 MEDLINE  
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[PMID]:29261729
[Au] Autor:Almeida Garcia R; Lima Pepino Macedo L; Cabral do Nascimento D; Gillet FX; Moreira-Pinto CE; Faheem M; Moreschi Basso AM; Mattar Silva MC; Grossi-de-Sa MF
[Ad] Endereço:Brasilia Federal University (UnB), Brasília - CEP, Brasília, Federal District, Brazil.
[Ti] Título:Nucleases as a barrier to gene silencing in the cotton boll weevil, Anthonomus grandis.
[So] Source:PLoS One;12(12):e0189600, 2017.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:RNA interference (RNAi) approaches have been applied as a biotechnological tool for controlling plant insect pests via selective gene down regulation. However, the inefficiency of RNAi mechanism in insects is associated with several barriers, including dsRNA delivery and uptake by the cell, dsRNA interaction with the cellular membrane receptor and dsRNA exposure to insect gut nucleases during feeding. The cotton boll weevil (Anthonomus grandis) is a coleopteran in which RNAi-mediated gene silencing does not function efficiently through dsRNA feeding, and the factors involved in the mechanism remain unknown. Herein, we identified three nucleases in the cotton boll weevil transcriptome denoted AgraNuc1, AgraNuc2, and AgraNuc3, and the influences of these nucleases on the gene silencing of A. grandis chitin synthase II (AgraChSII) were evaluated through oral dsRNA feeding trials. A phylogenetic analysis showed that all three nucleases share high similarity with the DNA/RNA non-specific endonuclease family of other insects. These nucleases were found to be mainly expressed in the posterior midgut region of the insect. Two days after nuclease RNAi-mediated gene silencing, dsRNA degradation by the gut juice was substantially reduced. Notably, after nucleases gene silencing, the orally delivered dsRNA against the AgraChSII gene resulted in improved gene silencing efficiency when compared to the control (non-silenced nucleases). The data presented here demonstrates that A. grandis midgut nucleases are effectively one of the main barriers to dsRNA delivery and emphasize the need to develop novel RNAi delivery strategies focusing on protecting the dsRNA from gut nucleases and enhancing its oral delivery and uptake to crop insect pests.
[Mh] Termos MeSH primário: Endonucleases/metabolismo
Inativação Gênica
Controle de Insetos/métodos
Gorgulhos/genética
[Mh] Termos MeSH secundário: Animais
Interferência de RNA
Reação em Cadeia da Polimerase em Tempo Real
Transcriptoma
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
EC 3.1.- (Endonucleases)
[Em] Mês de entrada:1801
[Cu] Atualização por classe:180116
[Lr] Data última revisão:
180116
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171221
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0189600



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