Transgênese e clonagem adotadas como biotécnicas reprodutivas em pequenos ruminantes / Transgenesis and cloning as reproductive biotechniques in small ruminants

Ovinos e caprinos são espécies que devido ao seu tamanho adequado, curto período gestacional e produção leiteira, tornaram-se importantes modelos na pesquisa pecuária, farmacêutica e biomédica. Assim, a engenharia genética tem sido amplamente aplicada à pesquisa com pequenos ruminantes. Caprinos e ovinos modificados geneticamente fornecem modelos valiosos para pesquisa sobre as funções dos genes, melhorando a exploração, obtendo produtos farmacêuticos no leite e aumentando a resistência às doenças. Além disso, o uso conjunto da clonagem e transgênese já foi responsável por marcos importantes na biotecnologia animal. Esta revisão destaca os avanços da clonagem e da engenharia genética nessas espécies obtidos nas últimas quatro décadas, com ênfase nas pesquisas realizadas no Brasil.

Sheep and goats are valuable species that due to their suitable size, short pregnancy period, and secretion of milk, have become important model in agricultural, pharmaceutical, and biomedical research. Thus, genome engineering has been widely applied to sheep and goat research. Gene-edited sheep and goats provide valuable models for investigations on gene functions, improving animal breeding, producing pharmaceuticals in milk and improving disease resistance. Additionally, the joint use of cloning and transgenesis has already been responsible for major milestones in animal biotechnology. This review highlights the advances of cloning and genome engineering in these species over the past four decades with particular emphasis on research in Brazil.

Lentivirus vectors fail to deliver transgenes into bovine zygotes after co-incubation with sperm during in vitro fertilization / [Lentivírus falham na entrega de transgenes em zigotos bovinos após coincubação com espermatozoide, durante fertilização in vitro]

As vantagens dos animais transgênicos têm sido demonstradas em diferentes aplicações, entretanto muitas metodologias usadas para gerar animais geneticamente modificados (GM) apresentam baixas taxas de eficiência. O objetivo deste estudo foi avaliar a entrega dos vetores lentivirais (VLs) em zigotos durante a fertilização in vitro (FIV), para gerar embriões GM, com o gene da proteína verde fluorescente (GFP) ou do fator IX de coagulação humana (FIX). Vetores lentivirais com os genes GFP (pLGW-GFP-LV) ou FIX (pLWE2-FIX-LV) foram utilizados na FIV ou na cultura de embriões in vitro (CIV). A coincubação de pLWE2-FIX-LV com espermatozoides e complexos oócitos-células do cumulus (COCs) durante a FIV diminuiu (P<0,05) as taxas de clivagem e de blastocistos, enquanto com pLGW-GFP-LV diminuiu (P<0,05) a taxa de blastocisto quando se comparou ao controle sem VL. A coincubação de pLWE2-FIX-LV e pLGW-GFP-LV com presumíveis zigotos durante a CIV não afetou (P>0,05) o desenvolvimento embrionário. A expressão da proteína GFP não foi detectada em embriões após a coincubação de FIV ou CIV, embora as células do cumulus expressassem a proteína até o dia oito de cultivo in vitro. Reações em cadeia da polimerase (PCR) não detectaram os genes GFP ou FIX em embriões, mas ambos foram detectados em células do cumulus. Assim, a coincubação de VL com espermatozoide bovino e COCs não é eficaz para produzir embriões geneticamente modificados por meio de FIV.(AU)

Parâmetros seminais de caprinos transgênicos para o fator estimulante de colônia de granulócitos humano / Seminal parameters of transgenic goats for the cologne stimulating factor of human granulocytes

Neste estudo, a avaliação do sêmen foi utilizada para investigar prováveis efeitos da presença do transgene sobre os parâmetros seminais de caprinos. Para tanto, amostras de sêmen foram colhidas semanalmente de três caprinos, sendo um não transgênico (NTG) e dois transgênicos (TG) para o Fator Estimulante de Colônia de Granulócitos humano (hG-CSF). Observou-se que, com exceção do volume ejaculado, não foram verificadas diferenças estatísticas para os dados relativos aos demais parâmetros: motilidade massal, concentração espermática, motilidade individual progressiva e patologias espermáticas. Concluiu-se que a presença do transgene hG-CSF não teve influência sobre os principais parâmetros seminais dos animais avaliados.

In this study, semen evaluation was used to investigate likely effects of transgene presence on seminal parameters of goats. For this purpose, semen samples were collected weekly from three goats, one non-transgenic (NTG) and two transgenic (TG) for the Human Granulocyte Colony Stimulating Factor (hG-CSF). It was observed that, with the exception of the ejaculated volume, no statistical differences were verified for the data related to the other parameters: mass motility, sperm concentration, individual progressive motility and spermatic pathologies. It was concluded that the presence of the hG-CSF transgene had no influence on the main seminal parameters of the animals evaluated.

Parâmetros seminais de caprinos transgênicos para o fator estimulante de colônia de granulócitos humano / Seminal parameters of transgenic goats for the cologne stimulating factor of human granulocytes

Neste estudo, a avaliação do sêmen foi utilizada para investigar prováveis efeitos da presença do transgene sobre os parâmetros seminais de caprinos. Para tanto, amostras de sêmen foram colhidas semanalmente de três caprinos, sendo um não transgênico (NTG) e dois transgênicos (TG) para o Fator Estimulante de Colônia de Granulócitos humano (hG-CSF). Observou-se que, com exceção do volume ejaculado, não foram verificadas diferenças estatísticas para os dados relativos aos demais parâmetros: motilidade massal, concentração espermática, motilidade individual progressiva e patologias espermáticas. Concluiu-se que a presença do transgene hG-CSF não teve influência sobre os principais parâmetros seminais dos animais avaliados.(AU)

In this study, semen evaluation was used to investigate likely effects of transgene presence on seminal parameters of goats. For this purpose, semen samples were collected weekly from three goats, one non-transgenic (NTG) and two transgenic (TG) for the Human Granulocyte Colony Stimulating Factor (hG-CSF). It was observed that, with the exception of the ejaculated volume, no statistical differences were verified for the data related to the other parameters: mass motility, sperm concentration, individual progressive motility and spermatic pathologies. It was concluded that the presence of the hG-CSF transgene had no influence on the main seminal parameters of the animals evaluated.(AU)

Fluido amniótico, alternativa eficiente para obtenção de células-tronco mesenquimais multipotentes / Amniotic fluid, efficient alternative to obtain multipotent mesenchymal stem cells

Na última década houve um crescente interesse na investigação quanto à presença de células-tronco no fluido amniótico, devido a facilidade de obtenção, isolamento e cultivo in vitro. Além disso, a utilização do fluido como fonte de células-tronco possibilita a obtenção de células com alto grau de indiferenciação e elevada taxa de proliferação in vitro e grande capacidade de diferenciação. Todas estas vantagens tornam o líquido amniótico uma fonte atrativa de células-tronco para utilização em biotecnologias reprodutivas, como a clonagem e transgênese, bem como ensaios clínicos e terapias celulares em animais para posterior utilização em seres humanos. Este artigo apresenta um panorama da pesquisa científica com células-tronco derivadas do fluido amniótico no mundo, a partir de levantamento bibliográfico de artigos científicos de pesquisadores brasileiros e estrangeiros.

In the last decade there has been a growing interest in investigation of presence of stem cells in amniotic fluid due to the ease of obtaining, isolating and in vitro culture. In addition, the use of amniotic fluid as a source of stem cells makes it possible to obtain cells with a high degree of indifferentiation and a high rate of in vitro proliferation and a great capacity for differentiation. All of these advantages make amniotic fluid an attractive source of stem cells for use in reproductive biotechnologies, such as cloning and transgenesis, as well as clinical trials in animals for further use in humans. This article presents a panorama of the scientific research with stem cells derived from amniotic fluid in the world, from a bibliographical survey of scientific articles of Brazilian and foreign researchers.

Fluido amniótico, alternativa eficiente para obtenção de células-tronco mesenquimais multipotentes / Amniotic fluid, efficient alternative to obtain multipotent mesenchymal stem cells

Na última década houve um crescente interesse na investigação quanto à presença de células-tronco no fluido amniótico, devido a facilidade de obtenção, isolamento e cultivo in vitro. Além disso, a utilização do fluido como fonte de células-tronco possibilita a obtenção de células com alto grau de indiferenciação e elevada taxa de proliferação in vitro e grande capacidade de diferenciação. Todas estas vantagens tornam o líquido amniótico uma fonte atrativa de células-tronco para utilização em biotecnologias reprodutivas, como a clonagem e transgênese, bem como ensaios clínicos e terapias celulares em animais para posterior utilização em seres humanos. Este artigo apresenta um panorama da pesquisa científica com células-tronco derivadas do fluido amniótico no mundo, a partir de levantamento bibliográfico de artigos científicos de pesquisadores brasileiros e estrangeiros.(AU)

In the last decade there has been a growing interest in investigation of presence of stem cells in amniotic fluid due to the ease of obtaining, isolating and in vitro culture. In addition, the use of amniotic fluid as a source of stem cells makes it possible to obtain cells with a high degree of indifferentiation and a high rate of in vitro proliferation and a great capacity for differentiation. All of these advantages make amniotic fluid an attractive source of stem cells for use in reproductive biotechnologies, such as cloning and transgenesis, as well as clinical trials in animals for further use in humans. This article presents a panorama of the scientific research with stem cells derived from amniotic fluid in the world, from a bibliographical survey of scientific articles of Brazilian and foreign researchers. (AU)

A espécie caprina como modelo para produção de proteínas recombinantes / Goat as a model for production of recombinant proteins

Vários sistemas para a produção de proteínas recombinantes têm sido utilizados para a otimização daprodução das mesmas. A glândula mamária é considerada um sistema muito interessante para a produção destasproteínas devido ao seu alto nível de expressão e à sua capacidade para realizar modificações pós-traducionais.Caprinos podem produzir elevadas quantidades de leite durante um longo período de lactação e, portanto,tornam-se importantes candidatos como biorreatores para a expressão de proteínas recombinantes no leite. Noentanto, caprinos transgênicos não são de fácil obtenção devido à baixa eficiência dos métodos e à expressãoimprevisível da proteína recombinante. O incremento na eficiência dos métodos na transgênese em caprinos éum grande desafio a superar. Esta revisão tem por objetivo apresentar o estado atual dos métodos tradicionais,bem como apresentar técnicas emergentes para obtenção eficiente de caprinos transgênicos.(AU)

Aiming the optimization for recombinant protein production, various systems have been used. Themammary gland is considered to be a very interesting system for the production of these proteins due to its highlevel of expression and its ability to perform post-translational modifications. Goats can produce largequantities of milk over a long lactation period, and therefore this species is an important candidate forrecombinant protein expression in milk. However, it is not easy to generate transgenic goats due to the lowefficiency of methods and unpredictable expression of recombinant protein. An increase in efficiency fortransgenic methodologies for goats is a big challenge to overcome. This review aims to present the state of art ofthe traditional methods, as well as the emerging technologies for obtaining transgenic goats efficiently.(AU)

A espécie caprina como modelo para produção de proteínas recombinantes / Goat as a model for production of recombinant proteins

Vários sistemas para a produção de proteínas recombinantes têm sido utilizados para a otimização daprodução das mesmas. A glândula mamária é considerada um sistema muito interessante para a produção destasproteínas devido ao seu alto nível de expressão e à sua capacidade para realizar modificações pós-traducionais.Caprinos podem produzir elevadas quantidades de leite durante um longo período de lactação e, portanto,tornam-se importantes candidatos como biorreatores para a expressão de proteínas recombinantes no leite. Noentanto, caprinos transgênicos não são de fácil obtenção devido à baixa eficiência dos métodos e à expressãoimprevisível da proteína recombinante. O incremento na eficiência dos métodos na transgênese em caprinos éum grande desafio a superar. Esta revisão tem por objetivo apresentar o estado atual dos métodos tradicionais,bem como apresentar técnicas emergentes para obtenção eficiente de caprinos transgênicos.

Aiming the optimization for recombinant protein production, various systems have been used. Themammary gland is considered to be a very interesting system for the production of these proteins due to its highlevel of expression and its ability to perform post-translational modifications. Goats can produce largequantities of milk over a long lactation period, and therefore this species is an important candidate forrecombinant protein expression in milk. However, it is not easy to generate transgenic goats due to the lowefficiency of methods and unpredictable expression of recombinant protein. An increase in efficiency fortransgenic methodologies for goats is a big challenge to overcome. This review aims to present the state of art ofthe traditional methods, as well as the emerging technologies for obtaining transgenic goats efficiently.

Transgenesis and gene edition in small ruminants / Transgênese e edição gênica em pequenos ruminantes

This review summarizes the main achievements with the use of transgenesis and genome editingtechnologies in sheep and goats. Transgenesis, also referred to as recombinant DNA (rDNA) technology, madepossible by the first time 30 years ago the addition of novel traits from a given species into a different one. Onthe other hand, more recently genome editing appears a much more precise method of making changes to thegenome of a plant, animal, or other living organism, allowing for the addition, substitution, or deletion ofspecific nucleotides in an organisms genome. With transgenesis, the introduction of new DNA into anorganisms genome was generally without control of the site of the genome in which the insertion of that rDNAconstruct would occur. With genome editing in contrast, researchers and developers of products can makespecific changes in precise locations of the genome. This concept was absolutely improved with the novelCRISPR/Cas system, making genome edition cheaper, more efficient, easier and affordable for every Laboratoryaround the world. This revolution that originally emerged from molecular biology and passed to biomedicine,has recently been applied to livestock and agriculture. In addition, the application of this technology in sheep,goats, pigs and cattle, also has been possible by the advance of assisted reproductive technologies for embryoproduction, micromanipulation, cryopreservation and transfer. In general, multidisciplinary approaches includingbasic research and technical improvements, participation of private actors and adequate regulation should bemerged to take advantage of this potent biotechnology in different countries.(AU)

Transgenesis and gene edition in small ruminants / Transgênese e edição gênica em pequenos ruminantes

This review summarizes the main achievements with the use of transgenesis and genome editingtechnologies in sheep and goats. Transgenesis, also referred to as recombinant DNA (rDNA) technology, madepossible by the first time 30 years ago the addition of novel traits from a given species into a different one. Onthe other hand, more recently genome editing appears a much more precise method of making changes to thegenome of a plant, animal, or other living organism, allowing for the addition, substitution, or deletion ofspecific nucleotides in an organism’s genome. With transgenesis, the introduction of new DNA into anorganism’s genome was generally without control of the site of the genome in which the insertion of that rDNAconstruct would occur. With genome editing in contrast, researchers and developers of products can makespecific changes in precise locations of the genome. This concept was absolutely improved with the novelCRISPR/Cas system, making genome edition cheaper, more efficient, easier and affordable for every Laboratoryaround the world. This revolution that originally emerged from molecular biology and passed to biomedicine,has recently been applied to livestock and agriculture. In addition, the application of this technology in sheep,goats, pigs and cattle, also has been possible by the advance of assisted reproductive technologies for embryoproduction, micromanipulation, cryopreservation and transfer. In general, multidisciplinary approaches includingbasic research and technical improvements, participation of private actors and adequate regulation should bemerged to take advantage of this potent biotechnology in different countries.

Aplicações da clonagem e transgênese em pequenos ruminantes / Cloning and transgenesis applied to small ruminants

Transgenic animals are those animals which are genetically modified. A foreign gene is inserted intothese animals to see certain characteristics. These animals can be helpful in producing organs, growthhormones and useful proteins for the humans. Transgenic farm animals were first developed in 1985 and wereuseful in the production of biopharmaceuticals shortly thereafter. The ideal transgenic animal produces plentyof milk, and has relatively short generation times. Thus, small ruminants have been selected for a number ofreasons, the chief of which is the short generational time of 18 months. Thus, this review aims to present thecharacteristics and main applications of transgenesis and cloning in small ruminants.(AU)

Aplicações da clonagem e transgênese em pequenos ruminantes / Cloning and transgenesis applied to small ruminants

Transgenic animals are those animals which are genetically modified. A foreign gene is inserted intothese animals to see certain characteristics. These animals can be helpful in producing organs, growthhormones and useful proteins for the humans. Transgenic farm animals were first developed in 1985 and wereuseful in the production of biopharmaceuticals shortly thereafter. The ideal transgenic animal produces plentyof milk, and has relatively short generation times. Thus, small ruminants have been selected for a number ofreasons, the chief of which is the short generational time of 18 months. Thus, this review aims to present thecharacteristics and main applications of transgenesis and cloning in small ruminants.

Perda gestacional em cabra transgênica para o hG-CSF / Pregnancy loss in transgenic goat for hG-CSF

The aim of this study was to report the occurrence of pregnancy loss in transgenic goat submitted toprenatal ultrasonography monitoring. We used on goat transgenic for human Granulocyte Colony StimulatingFactor (hG-CSF). The animal was submitted to mating after estrous synchronization. Ultrasound examinationswere carried out until D60 of pregnancy by transrectal via and after D60 by transabdominal via. Someparameters were observed such as morphology, organogenesis and formation of skeletal fetuses, viability withcardiac activity and fetus movements. During all sonographic evaluations, the conceptus showed normaldevelopment of their morphology and viability. Goat had fetal development completed by D150, however, therewere problems at birth and the fetus has died. In conclusion, the transgenic fetus remained viable and showednormal development during pregnancy, although still birth.(AU)

Perda gestacional em cabra transgênica para o hG-CSF / Pregnancy loss in transgenic goat for hG-CSF

The aim of this study was to report the occurrence of pregnancy loss in transgenic goat submitted toprenatal ultrasonography monitoring. We used on goat transgenic for human Granulocyte Colony StimulatingFactor (hG-CSF). The animal was submitted to mating after estrous synchronization. Ultrasound examinationswere carried out until D60 of pregnancy by transrectal via and after D60 by transabdominal via. Someparameters were observed such as morphology, organogenesis and formation of skeletal fetuses, viability withcardiac activity and fetus movements. During all sonographic evaluations, the conceptus showed normaldevelopment of their morphology and viability. Goat had fetal development completed by D150, however, therewere problems at birth and the fetus has died. In conclusion, the transgenic fetus remained viable and showednormal development during pregnancy, although still birth.

Ultrasonographic findings of the mammary gland, liver, gallbladder, spleen, and kidneys in transgenic goats for hG-CSF during induced lactation / Achados ultrassonográficos de glândula mamária, fígado, vesícula biliar, baço e rins de caprinos transgênicos para o hG-CSF durante lactação induzida

In transgenic murine models, the study of certain organs or tissues can be performed after euthanasia of some specimens. However, this practice may not be economically feasible when applied to livestock such as transgenic goats. It is necessary to use minimally invasive methods to perform in vivo studies of organs that may be affected by disorders related to the activity of the transgene, particularly during milk production, when the recombinant protein is secreted. The aim of this study was to describe ultrasonographic findings of the liver, gallbladder, spleen, kidneys, and mammary glands in transgenic goats for evaluating the effect of human granulocyte-colony stimulating factor (hG-CSF) expression in milk during induced lactation. Six female Canindé goats-three transgenic (T) and three non-transgenic (NT)-were subjected to hormone therapy to induce lactation; ultrasonographic examinations of the liver, gallbladder, spleen, kidneys, and mammary gland were performed during both the hormonal therapy and the lactation period at different intervals depending on the organ being examined. On Day 16 (Day 1 = hormonal therapy initiation), all goats were lactating and presented healthy mammary glands, characterized by echogenic parenchyma showing a granular echotexture. Transgenic and nontransgenic goats were compared on the basis of measurements and ultrasound images obtained from each organ. No differences between T and NT animals were observed in the examined area for the liver, gallbladder, spleen, and kidneys. Liver and renal echogenicity and appearance of gallbladder and portal and hepatic veins were similar in all females. Ultrasonographic findings of the mammary gland, liver, gallbladder, spleen, and kidneys in transgenic goats did not show any difference from those in nontransgenic goats.[...]

Em camundongos transgênicos, o estudo de certos órgãos ou tecidos pode ser viabilizado por meio da eutanásia de alguns exemplares. Entretanto, esta prática pode não ser economicamente viável quando aplicada a animais de produção, como caprinos transgênicos. É necessário utilizar métodos minimamente invasivos para estudar in vivo os órgãos que podem estar envolvidos em desordens relacionadas à atividade do transgene, principalmente durante a produção de leite, quando a proteína recombinante é secretada. O objetivo deste estudo foi descrever as características ultrassonográficas de fígado, vesícula biliar, baço, rins e glândula mamária de caprinos transgênicos expressando o fator estimulante de colônias de granulócitos humano (hG-CSF) no leite, durante lactação induzida. Seis fêmeas caprinas da raça Canindé: três não transgênicas (NT) e três transgênicas (T) foram submetidas a tratamento hormonal para indução da lactação; os exames ultrassonográficos de fígado, vesícula biliar, baço, rins e glândula mamária foram realizados durante tratamento hormonal e lactação, em diferentes intervalos de acordo com o órgão. No dia 16 (Dia 1 = início do tratamento hormonal) todas as cabras estavam em lactação e apresentando uma glândula mamária saudável, caracterizada por parênquima ecogênico, com uma ecotextura granular. As cabras T e NT foram comparadas em relação às mensurações e aparência ultrassonográfica obtida de cada órgão. Não foram observadas diferenças com relação à área para exame de fígado, vesícula biliar, baço e rins entre T e NT. A ecogenicidade de fígado e rins, a aparência da vesícula biliar e das veias porta e hepática foram similares em todas as fêmeas. As mensurações ultrassonográficas dos órgãos abdominais examinados não revelaram diferenças entre animais T e NT.[...]

Ultrasonographic findings of the mammary gland, liver, gallbladder, spleen, and kidneys in transgenic goats for hG-CSF during induced lactation / Achados ultrassonográficos de glândula mamária, fígado, vesícula biliar, baço e rins de caprinos transgênicos para o hG-CSF durante lactação induzida

In transgenic murine models, the study of certain organs or tissues can be performed after euthanasia of some specimens. However, this practice may not be economically feasible when applied to livestock such as transgenic goats. It is necessary to use minimally invasive methods to perform in vivo studies of organs that may be affected by disorders related to the activity of the transgene, particularly during milk production, when the recombinant protein is secreted. The aim of this study was to describe ultrasonographic findings of the liver, gallbladder, spleen, kidneys, and mammary glands in transgenic goats for evaluating the effect of human granulocyte-colony stimulating factor (hG-CSF) expression in milk during induced lactation. Six female Canindé goats-three transgenic (T) and three non-transgenic (NT)-were subjected to hormone therapy to induce lactation; ultrasonographic examinations of the liver, gallbladder, spleen, kidneys, and mammary gland were performed during both the hormonal therapy and the lactation period at different intervals depending on the organ being examined. On Day 16 (Day 1 = hormonal therapy initiation), all goats were lactating and presented healthy mammary glands, characterized by echogenic parenchyma showing a granular echotexture. Transgenic and nontransgenic goats were compared on the basis of measurements and ultrasound images obtained from each organ. No differences between T and NT animals were observed in the examined area for the liver, gallbladder, spleen, and kidneys. Liver and renal echogenicity and appearance of gallbladder and portal and hepatic veins were similar in all females. Ultrasonographic findings of the mammary gland, liver, gallbladder, spleen, and kidneys in transgenic goats did not show any difference from those in nontransgenic goats.[...](AU)

Em camundongos transgênicos, o estudo de certos órgãos ou tecidos pode ser viabilizado por meio da eutanásia de alguns exemplares. Entretanto, esta prática pode não ser economicamente viável quando aplicada a animais de produção, como caprinos transgênicos. É necessário utilizar métodos minimamente invasivos para estudar in vivo os órgãos que podem estar envolvidos em desordens relacionadas à atividade do transgene, principalmente durante a produção de leite, quando a proteína recombinante é secretada. O objetivo deste estudo foi descrever as características ultrassonográficas de fígado, vesícula biliar, baço, rins e glândula mamária de caprinos transgênicos expressando o fator estimulante de colônias de granulócitos humano (hG-CSF) no leite, durante lactação induzida. Seis fêmeas caprinas da raça Canindé: três não transgênicas (NT) e três transgênicas (T) foram submetidas a tratamento hormonal para indução da lactação; os exames ultrassonográficos de fígado, vesícula biliar, baço, rins e glândula mamária foram realizados durante tratamento hormonal e lactação, em diferentes intervalos de acordo com o órgão. No dia 16 (Dia 1 = início do tratamento hormonal) todas as cabras estavam em lactação e apresentando uma glândula mamária saudável, caracterizada por parênquima ecogênico, com uma ecotextura granular. As cabras T e NT foram comparadas em relação às mensurações e aparência ultrassonográfica obtida de cada órgão. Não foram observadas diferenças com relação à área para exame de fígado, vesícula biliar, baço e rins entre T e NT. A ecogenicidade de fígado e rins, a aparência da vesícula biliar e das veias porta e hepática foram similares em todas as fêmeas. As mensurações ultrassonográficas dos órgãos abdominais examinados não revelaram diferenças entre animais T e NT.[...](AU)

Production of recombinant therapeutic proteins in genetically engineered animals: the dawn of a new era / Produção de proteínas terapêuticas recombinantes em animais geneticamente modificados: o surgimento de uma nova era

Background: The production of transgenic animals has been envisioned as a viable strategy to improve food quality, animal yield, and for the production of bioproducts that can be used for the benefit of the human and animal population. Transgenic animals have been used to improve production traits, to add value to animal products, to minimize the impact on the environment, to promote disease resistance, and most notably, to produce recombinant proteins in natural fluids, such as milk, that can be collected, purified and used as biomedical products (biopharming). This review aims to discuss past and recent technological advances in animal transgenesis, and the perspective for biopharming in Brazil.Review: Since the production of recombinant human insulin from Escherichia coli in the 1970s, continuous development of new platforms has allowed a significant expansion in the biopharmaceutical market. The animal platform has been shown to be highly competitive by adding value as low cost implementation, production and scale up, as well as high productivity of synthesized proteins. The expression of recombinant proteins in milk represents the most developed system for production of biopharmaceutical drugs in animals, with two approved biopharmaceuticals for human use: Atryn®, a recombinant antithrombin produced in the milk of goats, approved in 2006 by European Medicines Agency (EMA) and in 2009 by US Food and Drug Administration (FDA), and more recently, Ruconest®, a recombinant human C1 esterase inhibitor protein (C1INH) produced in the milk of rabbits, first approved by EMA in 2012, followed by the FDA approval in 2014. Transgenic animals have been produced by many strategies that have gradually evolved over the decades, including the use of embryo microinjection, viral vectors and transposable elements, sperm-mediated gene transfer, and cloning by somatic cell nuclear transfer (SCNT).[...](AU)

Production of recombinant therapeutic proteins in genetically engineered animals: the dawn of a new era / Produção de proteínas terapêuticas recombinantes em animais geneticamente modificados: o surgimento de uma nova era

Background: The production of transgenic animals has been envisioned as a viable strategy to improve food quality, animal yield, and for the production of bioproducts that can be used for the benefit of the human and animal population. Transgenic animals have been used to improve production traits, to add value to animal products, to minimize the impact on the environment, to promote disease resistance, and most notably, to produce recombinant proteins in natural fluids, such as milk, that can be collected, purified and used as biomedical products (biopharming). This review aims to discuss past and recent technological advances in animal transgenesis, and the perspective for biopharming in Brazil.Review: Since the production of recombinant human insulin from Escherichia coli in the 1970s, continuous development of new platforms has allowed a significant expansion in the biopharmaceutical market. The animal platform has been shown to be highly competitive by adding value as low cost implementation, production and scale up, as well as high productivity of synthesized proteins. The expression of recombinant proteins in milk represents the most developed system for production of biopharmaceutical drugs in animals, with two approved biopharmaceuticals for human use: Atryn®, a recombinant antithrombin produced in the milk of goats, approved in 2006 by European Medicines Agency (EMA) and in 2009 by US Food and Drug Administration (FDA), and more recently, Ruconest®, a recombinant human C1 esterase inhibitor protein (C1INH) produced in the milk of rabbits, first approved by EMA in 2012, followed by the FDA approval in 2014. Transgenic animals have been produced by many strategies that have gradually evolved over the decades, including the use of embryo microinjection, viral vectors and transposable elements, sperm-mediated gene transfer, and cloning by somatic cell nuclear transfer (SCNT).[...]

Transgênese em caprinos

Entre todos os mamíferos transgênicos produzidos até hoje, a espécie caprina tem representado um excelente modelo em transgênese quando são considerados os fatores como a demanda do mercado para a proteína, o volume de leite produzido por lactação e as taxas reprodutivas. Várias proteínas recombinantes foram obtidas a partir de cabras transgênicas, e entre estas, a antitrombina humana, foi a primeira proteína recombinante de origem animal a ser aprovada como um medicamento para a utilização clínica em seres humanos. Esta revisão tem por objetivo apresentar o estado-da-arte em caprinos transgênicos dando especial ênfase aos resultados obtidos por nosso grupo.

Transgênese em caprinos / Transgenesis in goats

Entre todos os mamíferos transgênicos produzidos até hoje, a espécie caprina tem representado um excelente modelo em transgênese quando são considerados os fatores como a demanda do mercado para a proteína, o volume de leite produzido por lactação e as taxas reprodutivas. Várias proteínas recombinantes foram obtidas a partir de cabras transgênicas, e entre estas, a antitrombina humana, foi a primeira proteína recombinante de origem animal a ser aprovada como um medicamento para a utilização clínica em seres humanos. Esta revisão tem por objetivo apresentar o estado-da-arte em caprinos transgênicos dando especial ênfase aos resultados obtidos por nosso grupo.

Among all the transgenic mammalians produced so far, goats have represented an excellent model of transgenesis when considering the factors such as the market demand for protein, volume of milk produced per lactation and reproductive rate. Various recombinant proteins have been obtained from the transgenic goats, and among these, human antithrombin was the first recombinant protein of animal origin to be released as a drug for the clinical use in humans. This review aims to present the state-of-the-art in transgenic goats with special emphasis on results obtained by our group.