Our goal was to target silencing of the Plum pox virus coat protein (PPV CP) gene independently expressed in plants. Clone C-2 is a transgenic plum expressing CP. We introduced and verified, in planta, the effects of the inverse repeat of CP sequence split by a hairpin (IRSH) that was characterized in the HoneySweet plum. The IRSH construct was driven by two CaMV35S promoter sequences flanking the CP sequence and had been introduced into C1738 plum. To determine if this structure was enough to induce silencing, cross-hybridization was made with the C1738 clone and the CP expressing but PPV-susceptible C2 clone. In total, 4 out of 63 clones were silenced. While introduction of the IRSH is reduced due to the heterozygous character in C1738 plum, the silencing induced by the IRSH PPV CP is robust. Extensive studies, in greenhouse containment, demonstrated that the genetic resource of C1738 clone can silence the CP production. In addition, these were verified through the virus transgene pyramiding in the BO70146 BlueByrd cv. plum that successfully produced resistant BlueByrd BO70146 × C1738 (HybC1738) hybrid plums.
- MeSH
- biotechnologie metody MeSH
- genetické inženýrství metody MeSH
- odolnost vůči nemocem * MeSH
- slivoň genetika virologie MeSH
- transgeny MeSH
- umlčování genů * MeSH
- virové plášťové proteiny genetika metabolismus MeSH
- virus šarky švestky genetika patogenita MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
According to Darwin's theory, endless evolution leads to a revolution. One such example is the Clustered Regularly Interspaced Palindromic Repeats (CRISPR)-Cas system, an adaptive immunity system in most archaea and many bacteria. Gene editing technology possesses a crucial potential to dramatically impact miscellaneous areas of life, and CRISPR-Cas represents the most suitable strategy. The system has ignited a revolution in the field of genetic engineering. The ease, precision, affordability of this system is akin to a Midas touch for researchers editing genomes. Undoubtedly, the applications of this system are endless. The CRISPR-Cas system is extensively employed in the treatment of infectious and genetic diseases, in metabolic disorders, in curing cancer, in developing sustainable methods for fuel production and chemicals, in improving the quality and quantity of food crops, and thus in catering to global food demands. Future applications of CRISPR-Cas will provide benefits for everyone and will save countless lives. The technology is evolving rapidly; therefore, an overview of continuous improvement is important. In this review, we aim to elucidate the current state of the CRISPR-Cas revolution in a tailor-made format from its discovery to exciting breakthroughs at the application level and further upcoming trends related to opportunities and challenges including ethical concerns.
- MeSH
- Archaea metabolismus MeSH
- Bacteria metabolismus MeSH
- CRISPR-Cas systémy * MeSH
- dějiny 20. století MeSH
- dějiny 21. století MeSH
- dobytek MeSH
- editace genu metody MeSH
- genetické inženýrství dějiny metody MeSH
- genom MeSH
- lidé MeSH
- sekvence CRISPR MeSH
- zemědělské plodiny genetika MeSH
- zvířata MeSH
- Check Tag
- dějiny 20. století MeSH
- dějiny 21. století MeSH
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- historické články MeSH
- přehledy MeSH
- MeSH
- genetické inženýrství metody MeSH
- geneticky modifikované organismy * genetika MeSH
- genová knihovna MeSH
- geny genetika MeSH
- interdisciplinární výzkum metody MeSH
- myši genetika MeSH
- výzkum MeSH
- Check Tag
- myši genetika MeSH
- Publikační typ
- novinové články MeSH
- Geografické názvy
- Česká republika MeSH
We report for the first time, a comparison of two approaches for artificially induced triploidy in zebrafish (Danio rerio) using cold shock and heat shock treatments. Of the two methods, heat shock treatment proved more effective with a triploid production rate of 100% in particular females. Subsequently, triploid zebrafish larvae were used as recipients for intraperitoneal transplantation of ovarian and testicular cells originating from vas:EGFP strain in order to verify their suitability for surrogate reproduction. Production of donor-derived sperm was achieved in 23% of testicular cell recipients and 16% of ovarian cell recipients, indicating the suitability of triploids as surrogate hosts for germ cell transplantation. Success of the transplantation was confirmed by positive GFP signal detected in gonads of dissected fish and stripped sperm. Germline transmission was confirmed by fertilization tests followed by PCR analysis of embryos with GFP specific primers. Reproductive success of germline chimera triploids evaluated as fertilization rate and progeny development was comparable to control groups.
- MeSH
- chov metody MeSH
- dánio pruhované genetika MeSH
- genetické inženýrství metody veterinární MeSH
- průtoková cytometrie MeSH
- teplota MeSH
- triploidie * MeSH
- zárodečné buňky transplantace MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- srovnávací studie MeSH
- MeSH
- biologická terapie etika metody MeSH
- cílená genová oprava etika metody MeSH
- CRISPR-Cas systémy genetika MeSH
- editace genu * etika metody MeSH
- fertilizace in vitro metody MeSH
- genetická terapie * etika metody MeSH
- genetické inženýrství etika metody MeSH
- HIV genetika MeSH
- lidé MeSH
- receptory CCR5 genetika MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- novinové články MeSH
Fakulta vojenského zdravotnictví Univerzity obrany, Hradec Králové Proteasy jsou skupina enzymů mikrobiálního, rostlinného i živočišného původu s nezastupitelným významem v základním metabolismu. Jedná se však i o materiál, který je vhodný k biotechnologickým aplikacím v širokém spektru oblastí jako je potravinový průmysl, výroba čistících prostředků a medicínské využití. Tento přehledový článek shrnuje základní informace o proteasach a nastiňuje příklady jejich použití.
Proteases are enzymes microbial, plant or animal origin with crucial importance in the basic metabolism. It is also a suitable material for biotechnological applications in a wide range of fields of interests like food industry, cleaning agents and medicine applications. This review paper surveys basic data about proteases and provides representative examples of their application.
- MeSH
- biomedicínské technologie metody trendy MeSH
- biotechnologie * metody trendy MeSH
- chemický průmysl metody trendy MeSH
- enzymy chemie metabolismus MeSH
- farmaceutický průmysl metody trendy MeSH
- genetické inženýrství metody trendy využití MeSH
- geneticky modifikované mikroorganismy enzymologie genetika metabolismus MeSH
- hydrolýza * účinky léků MeSH
- lidé MeSH
- peptidy chemie metabolismus MeSH
- potravinářský průmysl metody trendy MeSH
- proteasy * chemie metabolismus MeSH
- proteiny chemie metabolismus MeSH
- statistika jako téma MeSH
- zemědělství metody trendy MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- přehledy MeSH
- Klíčová slova
- luspatercept,
- MeSH
- anemie * terapie MeSH
- chimerické antigenní receptory terapeutické užití MeSH
- genetické inženýrství metody MeSH
- imunoglobuliny - Fc fragmenty terapeutické užití MeSH
- kongresy jako téma MeSH
- lidé MeSH
- rekombinantní fúzní proteiny terapeutické užití MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- zprávy MeSH
Lactococcus lactis is a food-grade lactic acid bacterium that is used in the dairy industry as a cell factory and as a host for recombinant protein expression. The nisin-controlled inducible expression (NICE) system is frequently applied in L. lactis; however new tools for its genetic modification are highly desirable. In this work NICE was adapted for dual protein expression. Plasmid pNZDual, that contains two nisin promoters and multiple cloning sites (MCSs), and pNZPolycist, that contains a single nisin promoter and two MCSs separated by the ribosome binding site, were constructed. Genes for the infrared fluorescent protein and for the human IgG-binding DARPin were cloned in all possible combinations to assess the protein yield. The dual promoter plasmid pNZDual enabled balanced expression of the two model proteins. It was exploited for the development of a single-plasmid inducible CRISPR-Cas9 system (pNZCRISPR) by using a nisin promoter, first to drive Cas9 expression and, secondly, to drive single guide RNA transcription. sgRNAs against htrA and ermR directed Cas9 against genomic or plasmid DNA and caused changes in bacterial growth and survival. Replacing Cas9 by dCas9 enabled CRISPR interference-mediated silencing of the upp gene. The present study introduces a new series of plasmids for advanced genetic modification of lactic acid bacterium L. lactis.
- MeSH
- antibakteriální látky farmakologie MeSH
- CRISPR-Cas systémy MeSH
- editace genu metody MeSH
- fermentace MeSH
- genetické inženýrství metody MeSH
- genom bakteriální * MeSH
- guide RNA, Kinetoplastida genetika metabolismus MeSH
- imunoglobulin G genetika metabolismus MeSH
- klonování DNA MeSH
- Lactococcus lactis genetika metabolismus MeSH
- lidé MeSH
- methyltransferasy genetika metabolismus MeSH
- nisin farmakologie MeSH
- plazmidy chemie metabolismus MeSH
- promotorové oblasti (genetika) účinky léků MeSH
- proteiny tepelného šoku genetika metabolismus MeSH
- regulace genové exprese u bakterií * MeSH
- rekombinantní fúzní proteiny genetika metabolismus MeSH
- transgeny * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
