Consulta: subjectFacets:"Homologous recombination"
Registros recuperados: 8
Data/hora: 16/06/2024 06:39:25

A simple negative selection method to identify adenovirus recombinants using colony PCR
Provedor de dados: 69
Autores: Zhao,Yongliang; Dong,Zongming; Li,Tengfei; Huang,Gang.
Background: The AdEasy system is a fast-track system for generating recombinant adenoviruses using the efficient homologous recombination machinery between shuttle and adenovirus backbone plasmids in Escherichia coli BJ5183 cells. The key step is homologous recombination in BJ5183 cells, which is driven by RecA activity. However, culture time is stringently limited to reduce the damage to recombinant plasmids by RecA activity. Therefore, rapid identification of recombinant adenoviruses within the limited time-period is critical. Results: We developed a simple negative selection method to identify recombinant adenoviruses using colony PCR, which improves the efficiency of adenovirus recombination screening and packaging. Conclusions: The negative selection...
Tipo: Journal article
Palavras-chave: Adenovirus; AdEasy; Colony PCR; Homologous recombination; RecA.
Ano: 2014
URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582014000100008

Reviewing the characteristics of BRCA and PALB2-related cancers in the precision medicine era
Provedor de dados: 74
Autores: Macedo,Gabriel S.; Alemar,Barbara; Ashton-Prolla,Patricia.
Abstract Germline mutations in BRCA1 and BRCA2 (BRCA) genes confer high risk of developing cancer, especially breast and ovarian tumors. Since the cloning of these tumor suppressor genes over two decades ago, a significant amount of research has been done. Most recently, monoallelic loss-of-function mutations in PALB2 have also been shown to increase the risk of breast cancer. The identification of BRCA1, BRCA2 and PALB2 as proteins involved in DNA double-strand break repair by homologous recombination and of the impact of complete loss of BRCA1 or BRCA2 within tumors have allowed the development of novel therapeutic approaches for patients with germline or somatic mutations in said genes. Despite the advances, especially in the clinical use of PARP...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: BRCA1; BRCA2; Homologous recombination; Cancer predisposition; PARP inhibitors.
Ano: 2019
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572019000200215

Persistent double strand break accumulation does not precede cell death in an Olaparib-sensitive BRCA-deficient colorectal cancer cell model
Provedor de dados: 74
Autores: Paviolo,Natalia Soledad; Vega,María Belén de la; Pansa,María Florencia; García,Iris Alejandra; Calzetta,Nicolás Luis; Soria,Gastón; Gottifredi,Vanesa.
Abstract The poly (adenosine diphosphate (ADP)-ribosyl) polymerase inhibitors (PARPi) selectively kill cancer cells with BRCA1 or BRCA2 (BRCA)-mutations. It has been proposed that cell death induction after PARPi depends on unrepaired double strand breaks (DSBs) that accumulate due to the homologous recombination deficiency of BRCA-mutated cells. Such accumulation of DSBs is inferred mainly from the high levels of DNA damage markers like phosphorylated histone H2AX. Herein, we developed a model of isogenic cell lines to show that depletion of BRCA causes PARPi-triggered cell death, replication stress (phosphorylated-H2AX and 53BP1 foci), and genomic instability. However, persistent DSBs accumulation was not detected under the same experimental conditions....
Tipo: Info:eu-repo/semantics/article
Palavras-chave: GammaH2AX; Alternative end joining; Non-homologous end joining; Homologous recombination; PARP.
Ano: 2020
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572020000200303

Role of RadA and DNA Polymerases in Recombination-Associated DNA Synthesis in Hyperthermophilic Archaea
Provedor de dados: 5
Autores: Hogrel, Gaelle; Lu, Yang; Alexandre, Nicolas; Bossé, Audrey; Dulermo, Remi; Ishino, Sonoko; Ishino, Yoshizumi; Flament, Didier.
Among the three domains of life, the process of homologous recombination (HR) plays a central role in the repair of double-strand DNA breaks and the restart of stalled replication forks. Curiously, main protein actors involved in the HR process appear to be essential for hyperthermophilic Archaea raising interesting questions about the role of HR in replication and repair strategies of those Archaea living in extreme conditions. One key actor of this process is the recombinase RadA, which allows the homologous strand search and provides a DNA substrate required for following DNA synthesis and restoring genetic information. DNA polymerase operation after the strand exchange step is unclear in Archaea. Working with Pyrococcus abyssi proteins, here we show...
Tipo: Text
Palavras-chave: Homologous recombination; Archaea; DNA polymerase; Recombinase; DNA repair.
Ano: 2020
URL: https://archimer.ifremer.fr/doc/00641/75274/75539.pdf

From Gene Targeting to Genome Editing: Transgenic animals applications and beyond
Provedor de dados: 42
Autores: ROCHA-MARTINS,MAURÍCIO; CAVALHEIRO,GABRIEL R.; MATOS-RODRIGUES,GABRIEL E.; MARTINS,RODRIGO A.P.
ABSTRACTGenome modification technologies are powerful tools for molecular biology and related areas. Advances in animal transgenesis and genome editing technologies during the past three decades allowed systematic interrogation of gene function that can help model how the genome influences cellular physiology. Genetic engineering via homologous recombination (HR) has been the standard method to modify genomic sequences. Nevertheless, nuclease-guided genome editing methods that were developed recently, such as ZFN, TALEN and CRISPR/Cas, opened new perspectives for biomedical research. Here, we present a brief historical perspective of genome modification methods, focusing on transgenic mice models. Moreover, we describe how new techniques were discovered...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Homologous recombination; DNA repair; Cre-LoxP; ZFN; TALEN; CRISPR/Cas9.
Ano: 2015
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0001-37652015000301323

Gene homozygosis and mitotic recombination induced by camptothecin and irinotecan in Aspergillus nidulans diploid cells
Provedor de dados: 42
Autores: ESQUISSATO,GIOVANA N.M.; SANT'ANNA,JULIANE R. DE; FRANCO,CLAUDINÉIA C.S.; ROSADA,LÚCIA J.; SANTOS,PAULA A.S.R. DOS; CASTRO-PRADO,MARIALBA A.A. DE.
Mitotic recombination is a process involved in carcinogenesis which can lead to genetic loss through the loss of heterozygosity. The recombinogenic potentials of two anticancer drugs topoisomerase I inhibitors, camptothecin (CPT) and irinotecan (CPT-11), were evaluated in the present study. The homozygotization assay, which assess the induction of mitotic recombination and gene homozygosis, as well as the heterozygous A757//UT448 diploid strain of Aspergillus nidulanswere employed. The three non-cytotoxic concentrations of CPT (3.5 ng mL−1, 10.5 ng mL−1 and 17.4 ng mL−1) were found to induce both mitotic recombination and gene homozygosis. CPT treatment produced three diploids homozygous, for nutritional and conidia color genes, and Homozygotization...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Anticancer drugs; Homozygotization assay; Homologous recombination; Secondary malignancies.
Ano: 2014
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0001-37652014000401703

Construction of yellow fever virus subgenomic replicons by yeast-based homologous recombination cloning technique
Provedor de dados: 42
Autores: Queiroz,Sabrina R.A.; Silva,Andréa N.M.R.; Santos,Jefferson J.S.; Marques Jr,Ernesto T.A.; Bertani,Giovani R.; Gil,Laura H.V.G.
RNA replicon derived from Flavivirus genome is a valuable tool for studying viral replication independent of virion assembly and maturation, besides being a great potencial for heterologous gene expression. In this study we described the construction of subgenomic replicons of yellow fever virus by yeast-based homologous recombination technique. The plasmid containing the yellow fever 17D strain replicon (pBSC-repYFV-17D), previously characterized, was handled to heterologous expression of the green fluorescent protein (repYFV-17D-GFP) and firefly luciferase (repYFV-17D-Luc) reporter genes. Both replicons were constructed by homologous recombination between the linearized vector pBSC-repYFV-17D and the PCR product containing homologous 25 nucleotides ends...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Cloning technique; Homologous recombination; Replicon; Reporter gene; Yellow fever virus.
Ano: 2013
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0001-37652013000100159

Construction of engineered murine Embryonic stem cells with conditional knockout of FGFR2 depending on Cre-loxP
Provedor de dados: 47
Autores: Wang,Jianmin; Song,Ruihua; Chen,Lei; Yin,Liangjun; Chen,Bo; Sun,Jing; Gou,Yuanbing; Zhao,Ling; Chen,Lin.
Objective: To investigate the functions of Fibroblast Growth Factor Receptor-2 (FGFR2) at different stages of cell differentiation. The engineered murine embryonic stem (ES) cells with conditional knockout of FGFR2 were developed depending on Cre-loxP. Methods: Cre-loxP system was used in a conditional targeting vector. The competent AM-1 bacteria, which expressed Cre-recombinase, was used to confirm the Cre-mediated deletion of the floxed exons 7 and 8 of FGFR2. The targeting vector was electroporated into the ES cells, and the transfected ES cells were screened with G418 and Ganciclovir. Finally, the ES clones with correct targeting events were identified by Southern Blot and PCR. Results: The targeting vector with conditional knockout of murine FGFR2...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Fibroblast growth factor receptor-2 (FGFR2); Conditional knockout; Homologous recombination; Cre; ES cells.
Ano: 2006
URL: http://www.scielo.org.ar/scielo.php?script=sci_arttext&pid=S0327-95452006000200002