Introduction Of New Genetic Material Research Articles

Germplasm Resources Evaluation of Cultured Largemouth Bass (Micropterus salmoides) in China Based on Whole Genome Resequencing.

Background: Largemouth bass (Micropterus salmoides), a valuable freshwater fish species, has experienced significant genetic decline in China due to prolonged domestic breeding and limited introduction of new genetic material. It is necessary to have a comprehensive understanding of the genetic status of largemouth bass populations in China. Method: In this study, we conducted population genetic analyses on nine cultured largemouth bass populations using whole genome resequencing. Results: A total of 3.23 Tb of clean bases were generated, with average Q20 and Q30 values of 98.17% and 94.25%, respectively, and 2,140,534 high-quality SNPs were obtained. Relatively high genetic diversity was observed across all populations. Combined with linkage disequilibrium (LD) patterns, the Wanlu (WL) population possessed the highest genetic diversity, and the Longyou (LY) population possessed the lowest genetic diversity. Additionally, population structure analyses, including pairwise F-statistics, phylogenetic trees, PCA, and admixture analysis, revealed significant genetic differentiation, particularly between the WL, LY, and other 7 populations, while also indicating the occurrence of a common admixture event. Finally, TreeMix inferred migration events from the WL to the Chuanlu (CL) population and from the Taiwan breeding population (TWL) to the Guanglu (GL) population. Conclusions: These findings provide a critical foundation for developing conservation and breeding strategies for largemouth bass in China.

Synthetic Biology and the Goals of Conservation

ABSTRACT The introduction of new genetic material into wild populations, using novel biotechnology, has the potential to fortify populations against existential threats, and, controversially, create wild genetically modified populations. The introduction of new genetic variation into populations, which will have an ongoing future in areas of conservation interest, complicates long-held values in conservation science and park management. I discuss and problematize, in light of genetic intervention, what I consider the three core goals of conservation science: biodiversity, ecosystem services, and wilderness. This uneasy relationship, however, does not forgo the use of such interventions. I argue there is a case for the application of this technology within some interpretations of the moral frameworks of wilderness and biodiversity, despite apparent conflict between these values and the technology, and the possibility that this technology's use displays some limitations of the ecosystem services framework. Ultimately, the advent of biotechnology assisted restoration highlights the need for us to reevaluate our ethical concepts of conservation at times of global environmental and technological change.

Time is running out: Microsatellite data predict the imminent extinction of the boreal lynx (Lynx lynx) in France

The Eurasian lynx reappeared in the French Jura Mountains, in the late 1970s, as the result of the dispersion of lynxes from the Carpathian region reintroduced in Switzerland in the early 1970s. Since then, the state of this population has remained poorly known. One estimate gives the number of individuals between 120 and 150. Opportunistic observation and monitoring suggest poor connectivity with the populations in Germany or in other parts of Switzerland. Forty years after its comeback in France, we could expect a higher number of individuals, but possible inbreeding depression as well as threats such as poaching and collisions with cars may explain the limitation of number of individuals observed and may lead to a new extinction of the specie in this region. We used 23 microsatellite markers and 78 DNA samples, collected between 2008 and 2020, to establish the genetic state of the French Jura Mountain lynx population. Our results show extremely low genetic diversity with an observed heterozygosity (Ho) of 0.36 and an effective population size (Ne) of 38.2. When comparing the source population (Carpathian), the inbreeding level is very worrying (Fe = 0.41). These results are among the lowest recorded for the Eurasian lynx. To avoid extinction, which we estimate will occur in approximately 30 years, we recommend the introduction of new genetic material. Although reintroduction is controversial, solutions such as the replacement of poached lynxes and the exchange of orphan lynxes between care centers are good initial actions for population strengthening. This offers a rapid intervention while studying solutions for more sustainable conservation

Genetic diversity in Amazonian Jundiá (Leiarius marmoratus) stocks using heterologous primers

With the rise of world fish farming, the national scenario is favorable for using native fish for intensive farming. Among the catfish, the Amazonian Jundiá (Leiarius marmoratus) is a robust candidate, easy to grow and with good organoleptic characteristics in its flesh. For productive success in captivity, it is necessary to consider some questions about the species, such as genetic variability, which must have an acceptable level in a breeding stock, in order to maintain a good diversity; this reduces losses due to inbreeding and low diversity. Therefore, the objective of this study was to characterize the genetic variability of commercial stocks of L. marmoratus from the State of Mato Grosso through microsatellite molecular markers. We analyzed 143 individuals from three stocks. The mean heterozygosity and the inbreeding coefficients observed were 0.060; 0.084; 0.141; and 0.539; 0.562; 0.514, respectively, for the stocks of Campo Verde, Juína, and Nova Mutum. The Deviation in the Hardy-Weinberg equilibrium was observed in most of the loci in the three populations. Considering the genetic differentiation, it is concluded that the three populations are very close genetically, which requires introduction of new genetic material in the stocks to enrich them genetically for a later reproductive program.

2021 clinical trials update: Innovations in hemophilia therapy.

Therapies engineered to prolong clotting factor protein circulation time, manipulate the balance of pro-coagulant and anti-coagulant proteins, or introduce new genetic material to enable endogenous factor protein production dominate the clinical trial landscape of hemophilia. The availability of clotting factor concentrates and the establishment of primary prophylaxis have dramatically improved health outcomes for hemophilia patients. But, the burden of hemostatic therapy remains significant, and many barriers to consistent longitudinal use of prophylaxis exist. Several types of emerging therapeutics including engineered factor concentrates, substitutive therapies, rebalancing therapies, and gene transfer/editing all aim to reduce the challenges of current hemophilia treatment. Emerging treatment options may reduce treatment frequency or need for intravenous administration. They may also introduce new challenges in laboratory assessment of hemostasis. These novel therapies must not introduce significant new health risks and continue to support similar or improved outcomes. The potential ramifications of rebalancing the coagulation cascade, particularly in a stress or inflammatory state, or introduction of new genetic material are not trivial. The focus of this review is to provide an overview of active and recently completed clinical trials as well as emerging preclinical data investigating new therapeutic possibilities for hemophilia patients and potentially other rare bleeding disorders.

Abstract LB-067: Cargocytes: A novel cell therapy platform to drive anti-tumor immunity

Abstract Adoptive cell therapy has emerged as a powerful new tool to treat cancer and other diseases. However, the clinical utility of many cell-based therapies is limited by inadequate production of therapeutic factors, production of unwanted factors, unwanted engraftment into the tumor, and poor controllability resulting in poor pharmacokinetics. Genetic and bioengineering strategies can overcome many limitations, but modification of genomic DNA or the introduction of new genetic material into therapeutic cells raises substantial safety concerns, ultimately making FDA approval difficult. To address these limitations, we developed a novel platform for cell-based therapies in which any cell type, including immortalized cells, can be extensively engineered and loaded with therapeutic cargo, and then rendered safe for patient administration through removal of the nucleus. Enucleated cells (Cargocytes) retain many desired biologic functions (endogenous and engineered), such as viability up to 72 hours, retention of cell surface markers/proteins, secretion of bioactive molecules, robust in vitro and in vivo chemotaxis, and delivery of a wide range of cancer-fighting cargos. Using this technology, we demonstrate that Cargocytes derived from hTERT-immortalized mesenchymal stem cells (hTERT MSCs) and engineered to express therapeutic levels of IL-12 can potently activate an anti-tumor immune response in a preclinical mouse model of triple negative breast cancer. hTERT MSC-derived Cargocytes robustly delivered IL-12 to the tumor with definable pharmacokinetics, activated known IL-12 biomarkers, and induced infiltration of tumor-fighting immune cells that inhibited tumor growth and improved animal survival. In addition, co-administration of Cargocytes secreting IL-12 and anti-PD-L1 antibodies further improved animal survival without notable adverse events. Collectively, our findings indicate that Cargocytes provide a new means to deliver powerful immunomodulatory biologics to malignant tumors in a safe and controllable manner. The genetic tractability and excellent safety profile of the Cargoycte platform enables bioengineering of a wide range of designer therapeutics derived from genetically modified cells, immortalized stem cells, and even cancer cells, each with improved clinically relevant functions. Citation Format: Richard L. Klemke, Huawei Wang, Bei Liu, Christina Alarcon, Felicia Watson. Cargocytes: A novel cell therapy platform to drive anti-tumor immunity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr LB-067.

Current progress in non-surgical embryo transfer with fresh and vitrified/warmed pig embryos

Embryo transfer (ET) should play a critical role in the pig industry because it allows the movement and introduction of new genetic material into a herd with minimal risk of disease transmission and reduced transportation costs. In addition, embryo movement could prevent the potential health and welfare problems associated with transporting live pigs. Although the first successful ET was reported more than 60 years ago, the commercial use of this procedure in pigs is still in its infancy. The surgical requirements for embryo collection and transfer and the difficulties with embryo cryopreservation have prevented its use in pigs, unlike other livestock. However, new methodologies have been developed in the past decade to enable successful non-surgical ET and embryo cryopreservation that could allow the commercial use of ET in the pig industry. This review focuses on the development of these technologies with emphasis on our own findings. Specifically, we discuss the basic aspects of a non-surgical deep-uterine ET procedure and describe several factors that affect its efficacy in the transfer of fresh and short-term cultured embryos. Finally, we conclude with a brief discussion on the use of this procedure with long-term stored embryos.

Вплив «прилиття крові» півнів зарубіжних кросів на господарсько-корисні ознаки м’ясо-яєчних курей вітчизняної селекції

The purpose. To enrich economically beneficial attributes of meat-egg hens of domestic gene pool at crossing with genetic material of foreign selection. Methods. Selection-and-genetic, zootechnical, biometric, systemic approach, theoretical analysis and synthesis. Results. As a result of introduction of new genetic material of auk of foreign highly productive meat crosses «Kobb-500» and «Ross-308» into gene pool of meat-egg hens of local population their adaptive and productive attributes were improved, the level of variability that created good preconditions for increase of genetic potential and combinational variability of auk of domestic selection was raised. Conclusions. Expediency and perspective of use in the breeding purposes is proved of cocks of final hybrid of foreign meat cross «Ross-308» at their prolong use for improving indices of alive mass and egg production of meat-egg hens of domestic gene pool.

Genetic diversity of tambaqui broodstocks in stock enhancement programs.

Natural populations of tambaqui (Colossoma macropomum) have significantly decreased in recent decades especially due to human extraction activities. So that the environmental impact may be reduced, the restocking of fish and increase in fish production are enhanced. Genetic evaluations using molecular markers are essential for this purpose. Current study evaluates the genetic variability of two tambaqui broodstocks used in restocking programs. Sixty-five samples (33 samples from broodstock A and 32 samples from broodstock B) were collected. DNA was extracted from caudal fin samples, with the amplification of four microsatellite loci: Cm1A11 (EU685307) Cm1C8 (EU685308) Cm1F4 (EU685311) and Cm1H8 (EU685315). Fourteen alleles in the stock of broodstock A were produced, five alleles for Cm1A11 locus (230, 255, 260, 270 and 276 bp), three alleles Cm1C8 (239, 260, and 273 bp), two alleles Cm1F4 (211 and 245 bp), four alleles for Cm1H8 (275, 290, 320 and 331 bp) and two unique alleles were found for Cm1A11 loci (alleles 270 and 276 bp) and Cm1H8 (alleles 275 and 331 bp). In broodstock B, ten alleles were produced, the same alleles of the first stock except for alleles 270 and 276 bp in Cm1A11 locus and 275 and 331 bp in Cm1H8 locus. Broodstock A revealed low frequency alleles in Cm1A11 loci, Cm1C8, Cm1F4 and Cm1H8, whereas broodstock B had no locus with low allelic frequency. Loci Cm1A11, Cm1C8 and Cm1H8 exhibited significant deficit of heterozygotes in both broodstocks, revealing changes in Hardy-Weinberg equilibrium. Genetic diversity between stocks was 0.1120, whilst genetic similarity was 0.894, with FST rate = 0.05, and Nm = 3.93, indicating gene flow between the two broodstocks. Results show that broodstocks are genetically closely related, with no great genetic variability. Strategies such as a previous genetic analysis of breeding with its marking, use of a large Ne crossing between the most genetically divergent specimens, and the introduction of new genetic material to broodstocks may maximize genetic diversity and minimize inbreeding within the next generation.

Genetic diversity of tambaqui broodstocks in stock enhancement programs

Natural populations of tambaqui (Colossoma macropomum) have significantly decreased in recent decades especially due to human extraction activities. So that the environmental impact may be reduced, the restocking of fish and increase in fish production are enhanced. Genetic evaluations using molecular markers are essential for this purpose. Current study evaluates the genetic variability of two tambaqui broodstocks used in restocking programs. Sixty-five samples (33 samples from broodstock A and 32 samples from broodstock B) were collected. DNA was extracted from caudal fin samples, with the amplification of four microsatellite loci: Cm1A11 (EU685307) Cm1C8 (EU685308) Cm1F4 (EU685311) and Cm1H8 (EU685315). Fourteen alleles in the stock of broodstock A were produced, five alleles for Cm1A11 locus (230, 255, 260, 270 and 276 bp), three alleles Cm1C8 (239, 260, and 273 bp), two alleles Cm1F4 (211 and 245 bp), four alleles for Cm1H8 (275, 290, 320 and 331 bp) and two unique alleles were found for Cm1A11 loci (alleles 270 and 276 bp) and Cm1H8 (alleles 275 and 331 bp). In broodstock B, ten alleles were produced, the same alleles of the first stock except for alleles 270 and 276 bp in Cm1A11 locus and 275 and 331 bp in Cm1H8 locus. Broodstock A revealed low frequency alleles in Cm1A11 loci, Cm1C8, Cm1F4 and Cm1H8, whereas broodstock B had no locus with low allelic frequency. Loci Cm1A11, Cm1C8 and Cm1H8 exhibited significant deficit of heterozygotes in both broodstocks, revealing changes in Hardy-Weinberg equilibrium. Genetic diversity between stocks was 0.1120, whilst genetic similarity was 0.894, with FST rate = 0.05, and Nm = 3.93, indicating gene flow between the two broodstocks. Results show that broodstocks are genetically closely related, with no great genetic variability. Strategies such as a previous genetic analysis of breeding with its marking, use of a large Ne crossing between the most genetically divergent specimens, and the introduction of new genetic material to broodstocks may maximize genetic diversity and minimize inbreeding within the next generation.

DNA as therapeutics; an update.

Human gene therapy is the introduction of new genetic material into the cells of an individual with the intention of producing a therapeutic benefit for the patient. Deoxyribonucleic acid and ribonucleic acid are used in gene therapy. Over time and with proper oversight, human gene therapy might become an effective weapon in modern medicine's arsenal to help fight diseases such as cancer, acquired immunodeficiency syndrome, diabetes, high blood pressure, coronary heart disease, peripheral vascular disease, neurodegenerative diseases, cystic fibrosis, hemophilia and other genetic disorders. Gene therapy trials in humans are of two types, somatic and germ line gene therapy. There are many ethical, social, and commercial issues raised by the prospects of treating patients whose consent is impossible to obtain. This review summarizes deoxyribonucleic acid-based therapeutics and gene transfer technologies for the diseases that are known to be genetic in origin. Deoxyribonucleic acid-based therapeutics includes plasmids, oligonucleotides for antisense and antigene applications, deoxyribonucleic acid aptamers and deoxyribonucleic acidzymes. This review also includes current status of gene therapy and recent developments in gene therapy research.

PREVENTION OF URETHRAL BLOCKAGE FOLLOWING SEMEN COLLECTION IN TWO SPECIES OF LEMUR, VARECIA VARIEGATA VARIEGATA AND LEMUR CATTA

Lemurs are a diverse group of primates comprised of five families, all of which are found only on Madagascar and the Comoro Islands. Of the 60 known species, 17 are endangered and 5 of these are considered critically endangered. The effects of inbreeding on population health and viability have been well described; though negative inbreeding effects can be ameliorated through the introduction of new genetic material. Introduction of new individuals into a population can be extremely challenging because of the highly social nature of lemurs. Semen collection in lemur species is notoriously challenging, as the ejaculate forms a coagulum. During normal breeding, the coagulum forms a copulatory plug in the female. However, this coagulum can present a life-threatening situation when retained in the urethra abnormally following electroejaculation. This study investigates the use of ascorbic acid in preventing urethral blockage in two lemur species during semen collection, demonstrates successful collection of semen by electroejaculation from two species of lemur during the breeding season, and discusses removal of urethral plugs subsequent to semen collection. Semen was collected successfully from all animals. Urethral plugs formed during each collection and were abnormally retained in 2/11 collections. Both plugs were successfully and immediately removed with the use of retropulsion through a urethral catheter. Although the results of this study are encouraging, more investigation is required to establish whether or not this procedure can be safely performed in the field.

Extinction of island endemics: it is not inbreeding depression

Extinction of island endemics: it is not inbreeding depression

Applications of ultrasonography in the reproductive management of Dux magnus gentis venteris saginati

Dux magnus gentis venteris saginati is considered to be a Scottish delicacy; however, depleting wild stocks have resulted in attempts to farm them. Selective breeding has been successful in modifying...

Appraisal of the consequences of the DDT‐induced bottleneck on the level and geographic distribution of neutral genetic variation in Canadian peregrine falcons,Falco peregrinus

Peregrine falcon populations underwent devastating declines in the mid-20th century due to the bioaccumulation of organochlorine contaminants, becoming essentially extirpated east of the Great Plains and significantly reduced elsewhere in North America. Extensive re-introduction programs and restrictions on pesticide use in Canada and the United States have returned many populations to predecline sizes. A proper population genetic appraisal of the consequences of this decline requires an appropriate context defined by (i) meaningful demographic entities; and (ii) suitable reference populations. Here we explore the validity of currently recognized subspecies designations using data from the mitochondrial control region and 11 polymorphic microsatellite loci taken from 184 contemporary individuals from across the breeding range, and compare patterns of population genetic structure with historical patterns inferred from 95 museum specimens. Of the three North American subspecies, the west coast marine subspecies Falco peregrinus pealei is well differentiated genetically in both time periods using nuclear loci. In contrast, the partitioning of continental Falco peregrinus anatum and arctic Falco peregrinus tundrius subspecies is not substantiated, as individuals from these subspecies are historically indistinguishable genetically. Bayesian clustering analyses demonstrate that contemporary genetic differentiation between these two subspecies is mainly due to changes within F. p. anatum (specifically the southern F. p. anatum populations). Despite expectations and a variety of tests, no genetic bottleneck signature is found in the identified populations; in fact, many contemporary indices of diversity are higher than historical values. These results are rationalized by the promptness of the recovery and the possible introduction of new genetic material.

Piglets born after non-surgical deep intrauterine transfer of vitrified blastocysts in gilts

The aims of this study were: (1) to evaluate the effect of the number of previous estrus of recipient gilts on effectiveness of intrauterine insertion of a flexible catheter designed for non-surgical deep intrauterine catheterization during diestrus in pigs; and (2) to determine the farrowing rate and the litter size after non-surgical deep intrauterine embryo transfer (ET) of porcine blastocysts vitrified by the open pulled straw (OPS) method. In experiment 1, 27 large white hyperprolific gilts (LWh) with 2–6 previous estrus were used. Intrauterine insertions of the flexible catheter were carried out at day 5.5–6 of the estrous cycle (D0 = onset of estrus). During insertions, no or only moderate reactions were observed in 88.9% of gilts and was not related ( P > 0.05) to the number of estrus prior to the insertion periods. The number of the estrus had a significant effect ( P < 0.05) on the difficulties found during the procedure. In the 100% of gilts with two estrus ( N = 6) it was not possible to insert the flexible catheter through the cervix. In gilts with three or more estrus, it was possible to pass the cervix and to progress along a uterine horn in 80.9% of the cases. In 86.7% of the gilts, the tip of the flexible catheter achieved the second or third quarter of the uterine horn. In experiment 2, following non-surgical deep intrauterine transfer of 20 vitrified/warmed blastocysts, 9 Meishan recipients (42.9%) farrowed an average of 5.4 ± 0.8 piglets (range 3–9) of which 0.6 ± 0.3 piglets (range 0–2) were born dead. In conclusion, this study shows that it is possible to obtain birth of piglets following non-surgical deep intrauterine embryo transfer (ET) of vitrified/warmed blastocysts. Non-surgical deep intrauterine ET and OPS vitrification methods are promising procedures to be used together for the introduction of new genetic material in a farm.

Therapeutic angiogenesis: a fantastic new adventure.

A significant proportion of patients with coronary artery disease have symptoms refractory to medical treatment, yet are unsuitable for conventional revascularization techniques, like percutaneous coronary intervention and coronary artery bypass surgery. Such patients are potential candidates for alternative forms of coronary revascularization, like therapeutic angiogenesis. This strategy is designed to promote the development of supplemental collateral blood vessels that will act as endogenous bypass conduits. Two major avenues for achieving therapeutic angiogenesis are currently under intense investigation: gene therapy (the introduction of new genetic material into somatic cells to synthesize proteins that are missing, defective, or desired for specific therapeutic purposes) and protein-based therapy (administration of the growth factors, instead of the genes encoding for the growth factors responsible for angiogenesis). This article provides a concise review of the "components" of gene and protein-based therapy, namely, the growth factors, the vector (for gene therapy), the route of delivery, the therapeutic target, the desired therapeutic effect, and quantifiable clinical end points for trials of angiogenesis. Based on preliminary studies, the authors believe that therapeutic angiogenesis represents a promising novel therapy for treatment of the ischemic heart. In the future, angiogenesis will likely be offered as an adjunct to conventional revascularization strategies in subsets of patients who are only "suboptimally" revascularized with conventional techniques, and might evolve into a stand-alone treatment for some patients with nonrevascularizable disease.

Gene therapy techniques for peripheral arterial disease.

Somatic gene therapy is the introduction of new genetic material into selective somatic cells with resulting therapeutic benefits. Vascular wall and, subsequently, cardiovascular diseases have become an interesting target for gene therapy studies. Arteries are an attractive target for gene therapy since vascular interventions, both open surgical and endovascular, are well suited for minimally invasive, easily monitored gene delivery. Promising therapeutic effects have been obtained in animal models in preventing post-angioplasty restenosis and vein graft thickening, as well as increasing blood flow and collateral development in ischemic limbs. First clinical trials suggest a beneficial effect of vascular endothelial growth factor in achieving therapeutic angiogenesis in chronic limb ischemia and the efficacy of decoy oligonucleotides to prevent infrainguinal vein graft stenosis. However, further studies are mandatory to clarify the safety issues, to develop better gene delivery vectors and delivery catheters, to improve transgene expression, as well as to find the most effective and safe treatment genes.

Totipotency or multipotentiality of cultured cells: Applications and progress

The stable introduction of new genetic material into breeding programmes for farm animal species requires germ line modification. One route by which to achieve this is by the combination of embryo manipulation and molecular biological techniques. In the mouse, genetic manipulation of embryonic stem (ES) cells in culture followed by the production of chimeric animals by blastocyst injection has proved a powerful tool for germ line manipulation. In contrast, the absence of proven ES lines in farm animal species has restricted similar progress, the production of transgenic animals being limited to gene addition using the technique of pronuclear injection. An alternative and desirable route to germ line modification in farm animals is to produce offspring by nuclear transfer from a cell line which can be maintained in culture. In sheep this has now been achieved with the birth of live lambs from an embryo derived, epithelial like cell line. The combination of this technology with that developed in ES cells for genetic modification will herald a new era in the germ line modification of farm animal species.

Anti-viral strategies.

A variety of viral diseases that are refractory to vaccination or therapy are potentially amenable for treatment by gene therapy. Human gene therapy can be defined as the introduction of new genetic material into cells of an individual with resulting therapeutic benefit to the individual (Morgan and Anderson, 1993). Gene therapy for viral diseases requires the introduction of anti-viral genes into cells, to prevent or inhibit viral gene expression or function and consequently limit viral replication and pathogenesis.