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Arq. Ciênc. Vet. Zool. UNIPAR (Online) ; 24(1, cont.): e2403, jan-jun. 2021. tab, graf
Article in Portuguese | ID: biblio-1252764


O Whatman FTA-Card® é um papel-filtro quimicamente tratado, destinado à coleta, transporte, armazenamento de amostras para posterior extração de ácidos nucléicos. A tecnologia FTA-Card® é utilizada para manter estável DNA e RNA em temperatura ambiente, podendo ser utilizados para fixação de uma ampla variedade de material orgânico ou tecidos. Foram realizados testes para certificar sua eficiência na conservação do material a ser analisado com o intuito de eliminar a cadeia fria de conservação, agilizando o processo e diminuindo os custos da execução de exames moleculares associados ao diagnóstico de patologias. Foram testadas oito amostras de felinos na forma de sangue total e soro, para a extração utilizou-se o kit Magazorb RNA Total mini-prep kit (Promega®, EUA), para o diagnóstico foi utilizada a técnica de PCR em tempo real para amplificar o gene CI2 de mamíferos, a fim de visualizar a eficácia na conservação de ácidos nucleicos. A utilização desse método torna possível que o material biológico seja enviado por serviços de transporte postais, reduzindo os custos e viabilizando diagnósticos provenientes de áreas mais remotas.(AU)

Whatman FTA-Card® is a chemically treated filter paper intended for the collection, transport, and storage of samples for later extraction of nucleic acids. FTA-Card® technology is used to keep DNA and RNA stable at room temperature and can be used to fix a wide variety of organic material or tissues. Tests were carried out to certify its efficiency in the conservation of the material to be analyzed in order to eliminate the cold conservation chain, speeding up the process and decreasing the costs of performing molecular tests associated with the diagnosis of pathologies. By using this method, biological material can be sent by postal transport services, reducing costs and making diagnoses from more remote areas feasible. Samples of feline specimens were tested in the form of whole blood and serum, using the Magazorb RNA Total mini-prep kit (Promega®, USA) for the extraction. Diagnosis was performed using real-time PCR technique to amplify the mammalian CI2 gene in order to visualize the effectiveness in conserving nucleic acids.(AU)

Whatman FTA-Card® es un papel de filtro tratado químicamente, destinado a la recogida, transporte, almacenamiento de muestras para su posterior extracción de ácidos nucleicos. La tecnología FTA-Card® se usa para mantener el ADN y el ARN estables a la temperatura ambiente y se puede usar para la fijación de una amplia variedad de materiales o tejidos orgánicos. Se realizaron pruebas para certificar su eficiencia en la conservación del material a analizar con el fin de eliminar la cadena de frío de conservación, agilizando el proceso y reduciendo los costos de realización de pruebas moleculares asociadas al diagnóstico de patologías. Se analizaron ocho muestras felinas en forma de sangre total y suero, para la extracción se utilizó el mini-prep kit Magazorb RNA Total (Promega®, USA), para el diagnóstico se utilizó la técnica de PCR en tiempo real para amplificar el CI2 de mamífero gen, con el fin de visualizar la efectividad en la conservación de ácidos nucleicos. El uso de ese método permite el envío de material biológico por los servicios de transporte postal, lo que reduce los costes y permite realizar diagnósticos desde zonas más remotas.(AU)

Biocompatible Materials , DNA , Materials , Real-Time Polymerase Chain Reaction
Electron J Biotechnol ; 49: 29-33, Jan. 2021. tab, ilus
Article in English | LILACS | ID: biblio-1291632


BACKGROUND: Agkistrodon acutus, a traditional Chinese medicine, clinically used in the treatment of rheumatism, tumor, and cardiovascular and cerebrovascular diseases. Due to the unique medicinal value and the difficulty of artificial breeding of Agkistrodon acutus, the supply of Agkistrodon acutus on the market exceeds the demand, and a large number of its adulterants are found on the market. In this study, the cytb gene sequences of Agkistrodon acutus and 9 snakes were compared and analyzed, specific primers were designed, and specific PCR methods were established to detect Agkistrodon acutus medicinal samples on the market. RESULTS: This method was successfully applied to distinguish the snake from other adulterated species, and tested 18 Agkistrodon acutus samples randomly purchased from six cities. Twelve samples were counterfeit and six were genuine. The standard reference material of Agkistrodon acutus was cloned by molecular cloning and sequencing, and the gene sequence difference with other species was significant. It shows that the region could be used as the fingerprint region of the target species. CONCLUSIONS: The proposed method can be used as a species-specific marker and can be highly distinguished from other adulterated snake species, which is helpful to effectively avoid the problem of false sale of Agkistrodon acutus.

Animals , Polymerase Chain Reaction/methods , Agkistrodon/genetics , Cytochromes b/genetics , Mitochondria/genetics , Snakes , Species Specificity , DNA/analysis , Cloning, Molecular , Medicine, Chinese Traditional
Article in Chinese | WPRIM | ID: wpr-879618


OBJECTIVE@#To summarize clinical manifestations and results of genetic testing in 12 children with Gitelman syndrome (GS).@*METHODS@#Clinical data of the children was collected. Whole exome sequencing(WES) was carried out to screen potential variants of genomic DNA. Candidate variants were verified by Sanger sequencing.@*RESULTS@#The patients have included 10 boys and 2 girls, whom were diagnosed at between 2.8 to 15.0 year old. Six patients were due to infections, 5 were due to short stature, and 1 was due to lower limb weakness. All patients were found to carry variants of SLC12A3 gene, which included 11 with compound heterozygous variants and 1 with homozygous variant. All of the 19 alleles of the SLC12A3 gene carried by the patients were delineated, which included 15 missense variants, 2 frameshift variants and 2 splice region variants. These variants were unreported previously, which included c.578_582dupCCACC (p.Asn195Profs*109), c.251C>T (p.Pro84Leu) and c.2843G>A (p.Trp948X).@*CONCLUSION@#The clinical symptoms of GS in children are atypical and often seen in older children. For children with occasional hypokalemia associated with growth failure, GS should be suspected. The majority of GS children carry two pathogenic variants of the SLC12A3 gene, mainly compound heterozygotes, among which p.Thr60Met is the most common one. The discovery of new variants has enriched the spectrum of SLC12A3 gene variants.

Adolescent , Child , Child, Preschool , DNA , Female , Genetic Testing , Gitelman Syndrome/genetics , Humans , Hypokalemia/genetics , Male , Solute Carrier Family 12, Member 3/genetics
Article in Chinese | WPRIM | ID: wpr-879614


OBJECTIVE@#To investigate the genetic polymorphisms of 21 non-combined DNA index system short tandem repeat (STR) loci in Hainan Li population.@*METHODS@#DNA samples from 339 unrelated healthy individuals of Li population from Hainan Province were extracted and amplified with fluorescence labeled multiplex PCR system. PCR products were electrophoresed on an ABI3130 Genetic Analyzer following the manufacturer's instructions. Allele designation was performed with a GeneMapper ID-X by comparison with the allele ladder provided by the corresponding kit.@*RESULTS@#A total of 173 alleles and 489 genotypes were observed for the 21 STR loci, respectively. The frequencies of alleles and genotypes were 0.0010-0.5434 and 0.0020-0.3274, respectively. The heterozygosity varied from 0.639 to 0.833. Discrimination power (DP) was 0.803-0.948, power of exclusion for trio-paternity was 0.416-0.584, power of exclusion for duo-paternity was 0.140-0.238, the polymorphism information content(PIC) was 0.57-0.81, respectively. The total discrimination power (TDP), cumulative probability of exclusion for trio-paternity testing(CPE-trio) and cumulative probability of exclusion for duo-paternity testing (CPE-duo) were 0.999 999 999 999 99, 0.999 999 883 211 752, and 0.987 266, respectively.@*CONCLUSION@#The 21 STR loci are highly polymorphic and informative in the studied population and can be employed as supplementary loci in duo-paternity testing or cases with variant circumstances.

Asian Continental Ancestry Group/genetics , China , DNA , Gene Frequency , Genetics, Population , Humans , Microsatellite Repeats/genetics , Polymorphism, Genetic
Neotrop. ichthyol ; 19(1): e200102, 2021. tab, graf, ilus, mapas
Article in English | ID: biblio-1154969


A new species of Hyphessobrycon belonging to the Hyphessobrycon heterorhabdus species-group from the lower rio Tapajós, state of Pará, Brazil, is described. The new species is allocated into the Hyphessobrycon heterorhabdus species-group due to its color pattern, composed by an anteriorly well-defined, horizontally elongated humeral blotch that becomes diffuse and blurred posteriorly, where it overlaps with a conspicuous midlateral dark stripe that becomes blurred towards the caudal peduncle and the presence, in living specimens, of a tricolored longitudinal pattern composed by a dorsal red or reddish longitudinal stripe, a middle iridescent, golden or silvery longitudinal stripe, and a more ventrally-lying longitudinal dark pattern composed by the humeral blotch and dark midlateral stripe. It can be distinguished from all other species of the group by possessing humeral blotch with a straight or slightly rounded ventral profile, lacking a ventral expansion present in all other species of the group. The new species is also distinguished from Hyphessobrycon heterorhabdus by a 9.6% genetic distance in the cytochrome c oxidase I gene. The little morphological distinction of the new species when compared with its most similar congener, H. heterorhabdus, indicates that the new species is one of the first truly cryptic fish species described from the Amazon basin.(AU)

Uma nova espécie de Hyphessobrycon pertencente ao grupo Hyphessobrycon heterorhabdus é descrita da região do baixo rio Tapajós, estado do Pará, Brasil. A nova espécie é incluída no grupo Hyphessobrycon heterorhabdus devido ao seu padrão de coloração, composto por uma mancha umeral alongada, anteriormente bem definida, que se torna difusa e borrada posteriormente, onde se sobrepõe a uma conspícua faixa escura médio-lateral que se torna borrada próxima ao pedúnculo caudal, e pela presença, em exemplares vivos, de um padrão longitudinal tricolor, composto por uma faixa longitudinal vermelha ou avermelhada dorsal, uma faixa média iridescente dourada ou prateada e, mais ventralmente, o padrão longitudinal escuro composto pela faixa escura médio-lateral e mancha umeral. A espécie pode ser distinguida das outras espécies pertencentes ao grupo por possuir uma mancha umeral com região ventral retilínea ou levemente arredondada, sem uma expansão ventral presente nas demais espécies do grupo. A espécie também se diferencia de Hyphessobrycon heterorhabdus por uma distância genética de 9,6% no gene citocromo c oxidase I. A sutil diferença morfológica da nova espécie quando comparada ao seu congênere mais similar, H. heterorhabdus, indica que a nova espécie é uma das primeiras espécies de peixes verdadeiramente crípticas descritas da Bacia Amazônica.(AU)

Animals , DNA , Cytochromes c , Biodiversity , Characidae , Amazonian Ecosystem
J. venom. anim. toxins incl. trop. dis ; 27: e20200183, 2021. graf
Article in English | ID: biblio-1287097


The COVID-19 pandemic brought attention to studies about viral infections and their impact on the cell machinery. SARS-CoV-2, for example, invades the host cells by ACE2 interaction and possibly hijacks the mitochondria. To better understand the disease and to propose novel treatments, crucial aspects of SARS-CoV-2 enrolment with host mitochondria must be studied. The replicative process of the virus leads to consequences in mitochondrial function, and cell metabolism. The hijacking of mitochondria, on the other hand, can drive the extrusion of mitochondrial DNA (mtDNA) to the cytosol. Extracellular mtDNA evoke robust proinflammatory responses once detected, that may act in different pathways, eliciting important immune responses. However, few receptors are validated and are able to detect and respond to mtDNA. In this review, we propose that the mtDNA and its detection might be important in the immune process generated by SARS-CoV-2 and that this mechanism might be important in the lung pathogenesis seen in clinical symptoms. Therefore, investigating the mtDNA receptors and their signaling pathways might provide important clues for therapeutic interventions.(AU)

DNA/analysis , Genes, Mitochondrial , COVID-19 , Cytokines
Chinese Journal of Biotechnology ; (12): 1120-1130, 2021.
Article in Chinese | WPRIM | ID: wpr-878618


As the demand for high-performance computing continues to grow, traditional computing models are facing unprecedented challenges. Among the many emerging computing technologies, DNA computing has attracted much attention due to its low energy consumption and parallelism. The DNA circuit, which is the basis for DNA computing, is an important technology for the regulation and processing of the molecular information. This review highlights the basic principles of DNA computing, summarizes the latest research progress, and concludes with a discussion of the challenges of DNA computing. Such integrated molecular computing systems are expected to be widely used in the fields of aerospace, information security and defense system.

Article in English | WPRIM | ID: wpr-880633


OBJECTIVES@#To explore the correlation between cytosine-phosphoric-guanylic (CpG) site of Septin 9 gene and colorectal cancer, and to develop a real-time PCR detection system in plasma in patients with colorectal cancer.@*METHODS@#The methylation of training samples was detected by high-throughput sequencing technology, and the sites highly consistent with the clinical information of colorectal cancer were identified. Then the detection system of real-time PCR was designed to analyze the consistency of plasma and tissue based on methylationa sensitive enzyme digestion. Finally, 100 clinical trials were conducted to evaluate the performance of the detection system with the methylation sensitive enzyme digestion-real-time PCR.@*RESULTS@#The highly consistent sites, which were selected by high-throughput sequencing from 71 training set samples, was the 38th CpG. Based on the detection region, the screened methylation sensitive enzymes were @*CONCLUSIONS@#The 38th CpG site of Septin 9 detected by the detection system of methylation sensitive enzyme digestion-real-time PCR can highly predict the occurrence of colorectal cancer with great clinical application value.

Colorectal Neoplasms/genetics , CpG Islands/genetics , DNA , DNA Methylation , Humans , Plasma/metabolism , Septins/metabolism
Braz. arch. biol. technol ; 64: e21210076, 2021. tab, graf
Article in English | LILACS | ID: biblio-1278438


Abstract Species of the subfamily Holocentrinae, family Holocentidae, commonly called, squirrelfishes, are widely distributed from tropical to warm temperate waters. In Egypt, no data are available on genetic and evolutionary relationships of the family Holocentridae. Therefore, the study of the genetic relationship among Holocentrids species is crucial for proper management and convenient strategies. The purpose of this study was to evaluate the genetic relationship among eight species belonging to the family Holocentridae from the Mediterranean Sea and the Red Sea in Egypt using DNA barcoding. Based on this molecular marker, a phylogenetic tree was constructed for the studied Holocentrids species. 12S rRNA sequences discovered that Sargocentron caudimaculatum was clustered as closest taxa to Sargocentron spiniferum, being a sister group to each other. Also, Sargocentron punctatissimum and Sargocentron macrosquamis were more related to each other and formed a sister group. Moreover, this study discusses the building of genetic relationship among Sargocentron spinosissimum and Sargocentron macrosquamis for the first time to the other studied Sargocentrons. DNA barcoding using 12S rRNA gene provided efficient DNA barcodes for all of the studied species. The constructed phylogenetic tree based on the employed molecular marker provided the update for the barcoded Holocentridae species evolution.

Animals , Phylogeny , Sciuridae , DNA , Genes, rRNA
Braz. j. med. biol. res ; 54(5): e10743, 2021. tab, graf
Article in English | LILACS | ID: biblio-1180738


Amphiphilic copolymers have a wide variety of medical and biotechnological applications, including DNA transfection in eukaryotic cells. Still, no polymer-primed transfection of prokaryotic cells has been described. The reversible addition-fragmentation chain transfer (RAFT) polymer synthesis technique and the reversible deactivation radical polymerization variants allow the design of polymers with well-controlled molar mass, morphology, and hydrophilicity/hydrophobicity ratios. RAFT was used to synthesize two amphiphilic copolymers containing different ratios of the amphiphilic poly[2-(dimethyl-amino) ethyl methacrylate] and the hydrophobic poly [methyl methacrylate]. These copolymers bound to pUC-19 DNA and successfully transfected non-competent Escherichia coli DH5α, with transformation efficiency in the range of 103 colony-forming units per µg of plasmid DNA. These results demonstrate prokaryote transformation using polymers with controlled amphiphilic/hydrophobic ratios.

Polymers , DNA/genetics , Bacteria , Transfection , Cations
Acta amaz ; 50(4): 335-338, out. - dez. 2020.
Article in English | LILACS | ID: biblio-1146378


Muitas árvores tropicais possuem dossel alto e folhas não facilmente acessíveis. O uso de tecido de um órgão mais acessível (câmbio) para extração de DNA pode ser uma alternativa para estudos moleculares. Nós adaptamos uma metodologia viável para extrair DNA genômico de tecido cambial coletado no campo para avaliação com PCR. Testamos três condições de armazenamento (dois tampões e sílica gel) e quatro períodos após a coleta. Utilizamos protocolos descritos anteriormente e os testamos em três espécies encontradas em florestas amazônicas e outros biomas: Anadenanthera peregrina var. peregrina, Cedrela fissilis e Ceiba speciosa. Nosso protocolo foi eficaz na obtenção de DNA adequado para sequenciamento e genotipagem de microssatélites. Recomendamos o uso de sílica para armazenamento de longo prazo e o tampão com ácido ascórbico para curto prazo. (AU)

Ascorbic Acid , DNA , Dithiothreitol
Gac. méd. boliv ; 43(2): 179-183, dic. 2020. ilus
Article in Spanish | LILACS | ID: biblio-1249981


El artículo se centra en la utilización de la nueva herramienta, CRISPR (repeticiones palindrómicas cortas agrupadas a intervalos regulares), la cual permite editar los genomas de los seres vivos de manera más precisa que otras técnicas; a lo largo del artículo se mencionan trabajos relacionados con la detención de la angiogénesis, cáncer, Sarcoma de Kaposi en inmunodeficiencias, Parkinson, regeneración y modificación genética en humanos, todas estas investigaciones tiene en común la utilización de la herramienta CRISPR. También se comenta las complicaciones éticas que conlleva utilizar esta tecnología en el ADN de células embrionarias humanas, que según diferentes criterios, podrían llevar a generar seres humanos “mejorados”, es decir no solo sin susceptibilidad a enfermedades degenerativas o incurables, sino también modificados en aspectos físicos que no necesariamente estarían ligados a alguna patología.

The article focuses on the use of the new tool, CRISPR (short palindromic repetitions grouped at regular intervals), which allows editing the genomes of living beings more accurately than other techniques; Throughout the article, works related to the arrest of angiogenesis, cancer, Kaposi’s sarcoma in immunodeficiencies, Parkinson’s, regeneration and genetic modification in humans are mentioned, all these investigations have in common the use of the CRISPR tool. You can also comment on the ethical complications that involve using this technology in the DNA of human embryonic cells, which according to different criteria, carry out improved human beings, that is not only without susceptibility to degenerative or incurable diseases, but also modified in physical aspects that are not linked to any pathology.

DNA , Clustered Regularly Interspaced Short Palindromic Repeats , Sarcoma, Kaposi , Cells , Genome , Genetics , Neoplasms
Iatreia ; 33(3): 262-272, jul.-set. 2020. tab, graf
Article in Spanish | LILACS | ID: biblio-1143077


RESUMEN Dentro del mundo de las ciencias biológicas la terapia génica ha sido un tema llamativo desde su aparición. El desarrollo de nuevas tecnologías y avances en el campo de la bioingeniería como las nucleasas de dedos de zinc (ZFN), las nucleasas tipo activadores de transcripción (TALEN) y las repeticiones palindrómicas cortas agrupadas y regularmente interespaciadas (CRISPR/Cas9), abrieron las puertas a un sinnúmero de posibilidades en biología, entre ellas, la edición del genoma. Esta última consiste en la modificación directa del genoma a través de la introducción o escisión de secuencias nucleotídicas dentro de la hebra de ADN. Hoy en día su aplicación es extensa, desde el campo de la agroindustria y el control de plagas hasta el ámbito clínico con la "corrección" de enfermedades mendelianas, modulación de receptores inmunológicos en enfermedades infecciosas, modificaciones genéticas en líneas germinales, entre muchos otros empleos. Sin embargo, desde su descubrimiento en 1987, el sistema CRIS-PR/Cas9 no ha estado exento de polémica en aspectos bioéticos, la adquisición de su patente e, incluso, en cuanto a su eficacia. A pesar de las dificultades e incertidumbre que han surgido, el futuro del sistema es prometedor dada su sencillez y versatilidad de uso.

SUMMARY In biological sciences, genetic therapy constitutes a "trend topic" since its beginning. Development of new technologies in bioengineering as zinc-finger nucleases (ZFN), Transcription activator-like effector nucleases (TALEN) and Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR - Cas9) opened doors to a countless number of possibilities in biology, as genetic edition. Last one consists in a direct genomic modification through nucleotide sequences "introduction" or "cleavage" on DNA strands. Nowadays, its application is wide, since agroindustrial and pest control technologies to clinical area, with correcting mendelian diseases, modulating immunological receptors on infectious diseases, genetic modification in germ cells, among others. Nevertheless, since it's discovered in 1987, CRISPR - Cas9 system has not been exempt from controversy in bioethical aspects, patent acquisition and even about effectiveness. Despite the difficulties and uncertainty that have arisen, the future of the system is promising for its simplicity and versatility.

Humans , Publishing , DNA , Gene Editing
Electron. j. biotechnol ; 46: 50-54, jul. 2020. tab, graf, ilus
Article in English | LILACS | ID: biblio-1223243


BACKGROUND: Pomegranate (Punica granatum L.), one of the most important tropical fruits in Azad Jammu and Kashmir regions of Pakistan, is highly valued for its nutrition and medicinal purposes. Although pomegranate is native to this region, the genetic diversity among wild pomegranate accessions is currently unknown. Such information would be vital for germplasm conservation and breeding efforts. In the current study, genetic diversity among forty-eight wild pomegranate accessions collected from different agro-ecological zones of Azad Jammu and Kashmir was assessed using 41 simple sequence repeat (SSR) markers. RESULTS: The markers revealed 303 alleles averaging 7.39 alleles per marker. Polymorphic information content ranged from 0.12 (PGCT093B) to 0.88 (Pom006), with a mean of 0.54. The average genetic distance (GD) across all genotypes was 0.52, and was lowest between Chattar Class and Thorar genotypes (GD = 0.27), but highest between Khun Bandway and Akhor Ban (GD = 0.74). A neighbor-joining dendrogram separated the genotypes into three major clusters, with further sub-clustering within each cluster. CONCLUSIONS: Overall, the results presented here show significant genetic diversity among wild pomegranate accessions in Azad Jammu and Kashmir region of Pakistan. These accessions present a valuable genetic resource to breeding and cultivar improvement programs within the region.

Genetic Variation , Pomegranate/genetics , Pakistan , DNA , Microsatellite Repeats , Alleles
Rev. Hosp. Ital. B. Aires (2004) ; 40(2): 63-75, jun. 2020. graf, ilus, tab
Article in Spanish | LILACS | ID: biblio-1102739


El objetivo de este artículo es proporcionar una guía que sirva para la interpretación y seguimiento de los esfuerzos que se están desarrollando en todo el mundo con el objetivo de obtener una vacuna que pueda generar inmunidad contra el nuevo coronavirus SARS-CoV-2 de 2019, el agente causante de la enfermedad por coronavirus denominada COVID-19. Cinco meses después de haber sido detectada la enfermedad, ya hay 102 vacunas en distintos estadios de desarrollo, registradas por la Organización Mundial de la Salud (OMS), correspondientes a 8 plataformas vacunales con diferentes estrategias, y todos los días aparecen nuevas. Esto representará un enorme desafío de organismos internacionales, para la evaluación, comparación y selección de aquellas que cumplan con los criterios regulatorios indispensables de seguridad y eficacia y que, por otro lado, puedan ser producidas en cantidades suficientes para abastecer la demanda mundial. (AU)

The objective of this article is to provide a guide to help the interpretation and monitoring the efforts that are being carried out worldwide to obtain a vaccine that will be able to generate immunity against the new 2019 SARS-CoV-2 coronavirus, the viral agent causes the disease named COVID-19. Five months after the disease was detected, there are already 102 vaccines at different stages of development, registered by World Health Organization (WHO), corresponding to 8 vaccination platforms base on different strategies, and every day new ones appear. This will represent a huge challenge for international organizations, to evaluate, compare and selects those that will meet the essential regulatory criteria of safety and efficacy and that, would be able to be produced in enough quantities to supply the worldwide demand. Key words: SARS-Cov-2 vaccine, vaccine platform, COVID-19 strategy, attenuated virus, viral vector, viral proteins, viral DNA, viral RNA, nucleic acids, viral like particles, WHO. (AU)

Humans , Male , Female , Coronavirus Infections/therapy , SARS Virus/immunology , Pneumonia, Viral/therapy , DNA/therapeutic use , RNA/therapeutic use , Vaccines/therapeutic use , Nucleic Acids/therapeutic use , Protein S/immunology , Coronavirus Infections/virology , SARS Virus/physiology , SARS Virus/genetics , Disease Vectors
Article in Spanish | LILACS, BDNPAR | ID: biblio-1293132


La función original de los sistemas CRISPR/Cas es destruir el DNA de virus bacterianos. Este sistema ha evolucionado para identificar y cortar secuencias de diferentes DNA de virus de DNA evitando la infección. En la célula, está compuesto de genes Cas que producen nucleasas guiadas por RNA capaces de cortar el DNA. Si el RNA guía encuentra DNA de un virus con el que se puede emparejar, recluta a la nucleasa Cas9 que lo corta. Este sistema es utilizado in vitro para editar genes basándose en la producción de rupturas de doble cadena y su posterior reparación. Actualmente existen varias plataformas para el diseño de RNAs guía, aunque también es posible realizarlo de forma manual. Los componentes del sistema son entregados a la célula mediante un plásmido o una ribonucleoproteína. En esta revisión nos centraremos en la función original de CRISPR/Cas en procariotas y en cómo los investigadores la han modificado para proporcionar nuevas técnicas de edición de genomas. Discutiremos sobre las ventajas de esta nueva técnica, las formas en que podemos utilizarla y algunas de las limitaciones que aún encontramos en su aplicación

The original function of CRISPR/Cas systems is to destroy the DNA of bacterial viruses. This system has evolved to identify sequences of different DNA viruses and cut them in order to avoid infection. In the cell, the system is made up of Cas genes which produce RNA-guided nucleases capable of cutting DNA. If the guide RNA finds viral DNA with which it can pair up, it recruits the Cas9 nuclease to cut it. This system is used in vitro for gene edition, relying on the production of double-strand breaks and their subsequent repair. Currently, there are several platforms for the design of the guide RNA, and it is also possible to design it manually. The components of the system can be delivered to the cell through a plasmid or through a ribonucleoprotein. In this review we will focus on the original function of CRISPR/Cas in prokaryotes, and in how researchers have modified it in order to provide new genome editing techniques. We will discuss the advantages of this new technique, the ways in which it can be used, and some of the limitations found in its application

CRISPR-Cas Systems , Gene Editing , DNA
Article in English | WPRIM | ID: wpr-876629


@#There are a number of novel prenatal cytoogenetic analysis tests for obstetricians and gynecologists on detecting aneuploidies. In the recent years, screening of pregnant patients with non-invasive prenatal testing (NIPT) is one. As the spread of genomic medicine and preventive obstetrics continue, it is prudent for obstetricians and gynecologists to accept and optimize new screening modalities, whenever available. Chromosomal abnormalities are common. Worldwide, one out of 150 live births may involve chromosomal abnormalities. The American College of Obstetrics and Gynecologists (ACOG) and American College of Medical Genetics recommend invasive and non ? invasive prenatal testing (NIPT)3. The invasive testing, however, carries risk for procedure ? related miscarriage. 4This favors NIPT which avoids the risk. The current state of NIPT in the Philippines, is it was only in January 2018, were a NIPT workshop was conducted by the Society of Maternal Fetal Medicine.6 First, due to the minimal studies on personalized and precision medicine on prenatal testing, hence the strong move to conduct this study. In an extensive literature search review in Herdin, a local database and archives of Philippine Obstetrics and Gynecology, none specified researches on non ? invasive prenatal testing. Second, in our country alone, there is no provision for national prenatal tests. In our institution, it was already introduced but with no uptake yet. Because of this gap, scantiness and non - uptake on NIPT locally, hence the conduct of this study. The study aimed to investigate on the obstetricians and gynecologists (OB-GYNs) knowledge, attitude towards and practices (KAP) about NIPT. Majority of the OBGYNs were knowledgeable, had positive attitude and were practicing NIPT. Strikingly, a fourth of the respondents were not comfortable in explaining NIPT. The researcher recommends that there is a need to conduct this study on a larger scale cross - sectional survey and multiple studies due to the paucity of data.

Pregnancy , Female , Prenatal Diagnosis , Genetic Testing , Mass Screening , DNA
Article in Chinese | WPRIM | ID: wpr-826364


To investigate the relationship of both DNA methylation level and methylenetrahydrofolate reductase(MTHFR)gene polymorphism with ankylosing spondylitis(AS). Totally 200 Chinese AS patients with HLA-B27(+)and 120 healthy controls were included from Hunan Province.All the cases were diagnosed according to the 1984 modified New York criteria for AS.The DNA methylation was examined by cytosine extension method,while the MTHFR gene C677T polymorphism was analyzed by the polymerase chain reaction(PCR)and restriction fragment length polymorphism(RFLP).The plasma homocysteine(Hcy)level was examined by enzyme-linked immunosorbent assay(ELISA),while the red blood folate level was analyzed by the specific immunoassays. The ratio of the T/T genotype mutation in the AS group was significantly higher than in the control group(17.0% 5.0%;=9.874, =0.002).The plasma homocysteine concentration of AS group was(18.71 ± 2.42)μmol/L,which was significantly higher than that in normal control group [(10.97 ± 2.93)μmol/L](=24.402, <0.001).The plasma Hcy concentration of the T/T genotype [(21.70±1.80)μmol/L] was significantly higher than that of the C/C genotype[(18.31±1.94)μmol/L](=12.088, =0.01)and the C/T genotype [(17.80±2.18)μmol/L](=6.496, =0.01)in the AS group.The DNA methylation level of the T/T genotype in AS group was significantly lower than that in normal control group(=5.655, <0.001)and also significantly lower than those of the C/C genotype(=11.514, <0.001)and the C/T genotype in AS group(=10.287, <0.001). In the Han population in Hunan Province,the C677T polymorphism of the MTHFR gene is associated with the onset of AS.The T/T mutation at position 677 of the MTHFR gene is an important influencing factor for hyperhcyemia in the AS patients.The T/T mutation at position 677 of the MTHFR gene is associated with genomic DNA hypomethylation.Thus,hypomethylation of DNA may be one of the pathogenic mechanisms of AS.

DNA , DNA Methylation , Genomics , Genotype , Humans , Methylenetetrahydrofolate Reductase (NADPH2) , Polymorphism, Genetic , Spondylitis, Ankylosing