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recombinant DNA technology

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https://www.readbyqxmd.com/read/29785042/inducible-high-efficiency-crispr-cas9-targeted-gene-editing-and-precision-base-editing-in-african-trypanosomes
#1
Eva Rico, Laura Jeacock, Julie Kovářová, David Horn
The Cas9 endonuclease can be programmed by guide RNA to introduce sequence-specific breaks in genomic DNA. Thus, Cas9-based approaches present a range of novel options for genome manipulation and precision editing. African trypanosomes are parasites that cause lethal human and animal diseases. They also serve as models for studies on eukaryotic biology, including 'divergent' biology. Genome modification, exploiting the native homologous recombination machinery, has been important for studies on trypanosomes but often requires multiple rounds of transfection using selectable markers that integrate at low efficiency...
May 21, 2018: Scientific Reports
https://www.readbyqxmd.com/read/29770347/precision-tagging-a-novel-seamless-protein-tagging-by-combinational-use-of-type-ii-and-type-iis-restriction-endonucleases
#2
Zhen Xu, Yan-Ning Rui, John P Hagan, Dong H Kim
Protein tagging is a powerful tool for performing comprehensive analyses of the biological functions of a protein of interest owing to the existence of a wide variety of tags. It becomes indispensable in some cases, such as in tracking protein dynamics in a live cell or adding a peptide epitope due to the lack of optimal antibodies. However, efficiently integrating an array of tags into the gene of interest remains a challenge. Traditional DNA recombinant technology based on type II restriction endonucleases renders protein tagging tedious and inefficient as well as the introduction of an unwanted junction sequence...
February 5, 2018: Bio-protocol
https://www.readbyqxmd.com/read/29754246/a-rapid-combinatorial-approach-to-assembling-synthetic-prokaryotic-and-eukaryotic-protein-expression-vectors
#3
Rebecca Mullinax, Sarah E Johns, Denise Rhodes, Vivian Zhang, Nancy McKinney, Katherine A Felts, Carsten P Carstens, Peter Sheffield
Vector construction and gene cloning are ubiquitous techniques essential to all fields of biological and medical research. They are the first steps in many endeavors leading to expressing proteins to understand gene function and regulation. However, they can often be rate-limiting, particularly in multi-gene studies, due to the time and effort required to assemble gene constructs and to identify the optimal constructs for protein expression.The SureVector system was developed to address this by enabling the rapid and reliable assembly of multiple DNA modules into a recombinant plasmid containing a gene-of-interest (GOI)...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29753709/production-of-polyclonal-antibodies-for-capsicum-chlorosis-virus-cacv-infecting-chilli-in-india-through-recombinant-nucleocapsid-protein-expression-and-its-application
#4
B D Haokip, D Alice, R Selvarajan, K Nagendran, L Rajendran, S K Manoranjitham, G Karthikeyan
Bud necrosis and chlorotic spots causing virus affecting chilli crop in Tamil Nadu (India) was identified as Capsicum chlorosis virus (CaCV). Specific primers were used for amplification and sequencing of the nucleocapsid protein (NP) gene. Polyclonal antibody against the bacterially expressed NP from the CaCV-TN-CBE isolate was produced using recombinant DNA technology. NP gene was subcloned into the pET-28a (+) vector and expressed by transformation in BL21 (DE3) pLysS. The expressed protein was about ~34 kDa and was confirmed through western blot analysis using Groundnut bud necrosis virus (GBNV) polyclonal antiserum from ICRISAT, India...
May 10, 2018: Journal of Virological Methods
https://www.readbyqxmd.com/read/29714581/crispr-cas9-a-precise-approach-to-genome-engineering
#5
Jorge E Simón, Ángel S Rodríguez, Nelson Santiago Vispo
In the last few decades, genomic manipulation has made significant progress as a result of the development of recombinant DNA technologies; however, more often than not, these techniques have been costly and labor intensive. In contrast, recently developed next-generation sequencing (NGS) technologies have provided a cheaper, faster, and easier process to study genomics. In particular, an NGS technique emerged from bacterial CRISPR-associated protein-9 nuclease (Cas9) as a revolutionary method to modify, regulate, or mark specific genomic sequences on virtually any organism...
January 1, 2018: Therapeutic Innovation & Regulatory Science
https://www.readbyqxmd.com/read/29710373/transforming-plant-biology-and-breeding-with-crispr-cas9-cas12-and-cas13
#6
REVIEW
Patrick Schindele, Felix Wolter, Holger Puchta
Currently, biology is revolutionized by ever growing applications of the CRISPR/Cas system. As discussed in this Review, new avenues have opened up for plant research and breeding by the use of the sequence-specific DNases Cas9 and Cas12 (formerly named Cpf1) and, more recently, the RNase Cas13 (formerly named C2c2). Although double strand break-induced gene editing based on error-prone non-homologous end joining has been applied to obtain new traits, such as powdery mildew resistance in wheat or improved pathogen resistance and increased yield in tomato, improved technologies based on CRISPR/Cas for programmed change of plant genomes via homologous recombination have recently been developed...
April 30, 2018: FEBS Letters
https://www.readbyqxmd.com/read/29691470/crispr-cas9-genome-editing-technology-significantly-accelerated-herpes-simplex-virus-research
#7
REVIEW
Dong Wang, Xian-Wang Wang, Xiao-Chun Peng, Ying Xiang, Shi-Bao Song, Ying-Ying Wang, Lin Chen, Victoria W Xin, Yan-Ning Lyu, Jiafu Ji, Zhao-Wu Ma, Cheng-Bin Li, Hong-Wu Xin
Herpes simplex viruses (HSVs) are important pathogens and ideal for gene therapy due to its large genome size. Previous researches on HSVs were hampered because the technology to construct recombinant HSVs were based on DNA homology-dependent repair (HDR) and plaque assay, which are inefficient, laborious, and time-consuming. Fortunately, clustered regularly interspaced short palindromic repeat/CRISPR-associated protein 9 (CRISPR/Cas9) recently provided the possibility to precisely, efficiently, and rapidly edit genomes and indeed is successfully being used in HSVs...
April 25, 2018: Cancer Gene Therapy
https://www.readbyqxmd.com/read/29687035/pharmacology-of-recombinant-adeno-associated-virus-production
#8
REVIEW
Magalie Penaud-Budloo, Achille François, Nathalie Clément, Eduard Ayuso
Recombinant adeno-associated viral (rAAV) vectors have been used in more than 150 clinical trials with a good safety profile and significant clinical benefit in many genetic diseases. In addition, due to their ability to infect non-dividing and dividing cells and to serve as efficient substrate for homologous recombination, rAAVs are being used as a tool for gene-editing approaches. However, manufacturing of these vectors at high quantities and fulfilling current good manufacturing practices (GMP) is still a challenge, and several technological platforms are competing for this niche...
March 16, 2018: Molecular Therapy. Methods & Clinical Development
https://www.readbyqxmd.com/read/29685820/mastering-complexity-towards-bottom-up-construction-of-multifunctional-eukaryotic-synthetic-cells
#9
REVIEW
Kerstin Göpfrich, Ilia Platzman, Joachim P Spatz
With the ultimate aim to construct a living cell, bottom-up synthetic biology strives to reconstitute cellular phenomena in vitro - disentangled from the complex environment of a cell. Recent work towards this ambitious goal has provided new insights into the mechanisms governing life. With the fast-growing library of functional modules for synthetic cells, their classification and integration become increasingly important. We discuss strategies to reverse-engineer and recombine functional parts for synthetic eukaryotes, mimicking the characteristics of nature's own prototype...
April 21, 2018: Trends in Biotechnology
https://www.readbyqxmd.com/read/29682272/maintenance-of-fish-health-in-aquaculture-review-of-epidemiological-approaches-for-prevention-and-control-of-infectious-disease-of-fish
#10
REVIEW
Ayalew Assefa, Fufa Abunna
Aquaculture is rapidly growing part of agriculture worldwide. It makes up around 44 percent of total fish production globally. This increased growth of production is achieved despite facing many challenges in the aquaculture environment. Among production limiting challenges, the infectious disease takes the lion share by causing multibillion-dollar loss annually. To reduce the impact of the fish disease, it is necessary to address health constraints based on scientifically proven and recommended ways. This review aims at pointing out some of the best approaches to prevention and control of infectious disease in aquaculture...
2018: Veterinary Medicine International
https://www.readbyqxmd.com/read/29680684/fungalbraid-a-goldenbraid-based-modular-cloning-platform-for-the-assembly-and-exchange-of-dna-elements-tailored-to-fungal-synthetic-biology
#11
Miguel Hernanz-Koers, Mónica Gandía, Sandra Garrigues, Paloma Manzanares, Lynne Yenush, Diego Orzaez, Jose F Marcos
Current challenges in the study and biotechnological exploitation of filamentous fungi are the optimization of DNA cloning and fungal genetic transformation beyond model fungi, the open exchange of ready-to-use and standardized genetic elements among the research community, and the availability of universal synthetic biology tools and rules. The GoldenBraid (GB) cloning framework is a Golden Gate-based DNA cloning system developed for plant synthetic biology through Agrobacterium tumefaciens-mediated genetic transformation (ATMT)...
April 19, 2018: Fungal Genetics and Biology: FG & B
https://www.readbyqxmd.com/read/29680240/peptide-derivatives-of-erythropoietin-in-the-treatment-of-neuroinflammation-and-neurodegeneration
#12
Ilkcan Ercan, Kemal Ugur Tufekci, Ezgi Karaca, Sermin Genc, Kursad Genc
During the past 35 years, recombinant DNA technology has allowed the production of a wide range of hematopoietic and neurotrophic growth factors including erythropoietin (EPO). These have emerged as promising protein drugs in various human diseases. Accumulated evidences have recently demonstrated the neuroprotective effect of EPO in preclinical models of acute and chronic degenerative disorders. Nevertheless, tissue protective effect of EPO could not be translated to the clinical trials because of common lethal thromboembolic events, erythropoiesis and hypertension...
2018: Advances in Protein Chemistry and Structural Biology
https://www.readbyqxmd.com/read/29680238/in-silico-tools-and-databases-for-designing-peptide-based-vaccine-and-drugs
#13
Salman Sadullah Usmani, Rajesh Kumar, Sherry Bhalla, Vinod Kumar, Gajendra P S Raghava
The prolonged conventional approaches of drug screening and vaccine designing prerequisite patience, vigorous effort, outrageous cost as well as additional manpower. Screening and experimentally validating thousands of molecules for a specific therapeutic property never proved to be an easy task. Similarly, traditional way of vaccination includes administration of either whole or attenuated pathogen, which raises toxicity and safety issues. Emergence of sequencing and recombinant DNA technology led to the epitope-based advanced vaccination concept, i...
2018: Advances in Protein Chemistry and Structural Biology
https://www.readbyqxmd.com/read/29658862/progress-in-gene-therapy-for-chronic-heart-failure
#14
Zhi-Qiang Yin, Wan-Hong Xing
Chronic heart failure (CHF) is still the leading cause of morbidity and mortality worldwide, and carries with it large economic and social burdens. Although steady and substantial progress has been made in reducing mortality from heart failure using conventional treatments, novel pharmacologic and surgical interventions have not been effective in extending five year survival rates. Therefore, it is necessary to explore new therapies. Gene therapy was introduced in 1970s with the development of recombinant DNA technology...
March 5, 2018: Heart Surgery Forum
https://www.readbyqxmd.com/read/29656078/microbial-expression-systems-for-membrane-proteins
#15
REVIEW
Marvin V Dilworth, Mathilde S Piel, Kim E Bettaney, Pikyee Ma, Ji Luo, David Sharples, David R Poyner, Stephane R Gross, Karine Moncoq, Peter J F Henderson, Bruno Miroux, Roslyn M Bill
Despite many high-profile successes, recombinant membrane protein production remains a technical challenge; it is still the case that many fewer membrane protein structures have been published than those of soluble proteins. However, progress is being made because empirical methods have been developed to produce the required quantity and quality of these challenging targets. This review focuses on the microbial expression systems that are a key source of recombinant prokaryotic and eukaryotic membrane proteins for structural studies...
April 12, 2018: Methods: a Companion to Methods in Enzymology
https://www.readbyqxmd.com/read/29626699/therapeutics-incorporating-blood-constituents
#16
REVIEW
Phapanin Charoenphol, Katie Oswalt, Corey J Bishop
Blood deficiency and dysfunctionality can result in adverse events, which can primarily be treated by transfusion of blood or the re-introduction of properly functioning sub-components. Blood constituents can be engineered on the sub-cellular (i.e., DNA recombinant technology) and cellular level (i.e., cellular hitchhiking for drug delivery) for supplementing and enhancing therapeutic efficacy, in addition to rectifying dysfunctioning mechanisms (i.e., clotting). Herein, we report the progress of blood-based therapeutics, with an emphasis on recent applications of blood transfusion, blood cell-based therapies and biomimetic carriers...
April 4, 2018: Acta Biomaterialia
https://www.readbyqxmd.com/read/29587653/efficient-anchoring-of-alien-chromosome-segments-introgressed-into-bread-wheat-by-new-leymus-racemosus-genome-based-markers
#17
Offiong Ukpong Edet, June-Sik Kim, Masanori Okamoto, Kousuke Hanada, Tomoyuki Takeda, Masahiro Kishii, Yasir Serag Alnor Gorafi, Hisashi Tsujimoto
BACKGROUND: The tertiary gene pool of bread wheat, to which Leymus racemosus belongs, has remained underutilized due to the current limited genomic resources of the species that constitute it. Continuous enrichment of public databases with useful information regarding these species is, therefore, needed to provide insights on their genome structures and aid successful utilization of their genes to develop improved wheat cultivars for effective management of environmental stresses. RESULTS: We generated de novo DNA and mRNA sequence information of L...
March 27, 2018: BMC Genetics
https://www.readbyqxmd.com/read/29573495/efficient-in-planta-gene-targeting-in-arabidopsis-using-egg-cell-specific-expression-of-the-cas9-nuclease-of-s-aureus
#18
Felix Wolter, Jeanette Klemm, Holger Puchta
Gene targeting (GT), the programmed change of genomic sequences by homologous recombination (HR), is still a major challenge in plants. We previously developed an in planta GT strategy by simultaneously releasing from the genome a dsDNA donor molecule and creating a DSB at a specific site within the targeted gene. Using Cas9 form S. pyogenes (SpCas9) under the control of a ubiquitin gene promoter, we obtained seeds harbouring GT events, although at a low frequency. In the present research we tested different developmentally controlled promotors and different kinds of DNA lesions for their ability to enhance GT of the acetolactate synthase (ALS) gene of Arabidopsis...
March 24, 2018: Plant Journal: for Cell and Molecular Biology
https://www.readbyqxmd.com/read/29562451/-an-experimental-study-of-cd4-targeted-chimeric-antigen-receptor-modified-t-cell-with-anti-lymphoma-activity
#19
G H Chen, H W Huang, Y Wang, H W Liu, L J Xu, X Ma, S L Xue, X F He, Y Wang, B Gu, C X Li, H Y Qiu, X W Tang, Z M Jin, M Miao, A N Sun, D P Wu
Objective: To study the specific killing effect of CD4 membrane protein targeted chimeric antigen receptor modified T (CAR-T) cell. Methods: The second generation CD4 targeted chimeric antigen receptor containing 4-1BB costimulation domain was insert into lentiviral vector through recombinant DNA technology. Lentivirus was prepared and packaged by 293T cells with four plasmids. Beads activated T cells were transduced with lentivirus and the transduction efficiency was checked with Protein L and flow cytometry...
February 14, 2018: Zhonghua Xue Ye Xue za Zhi, Zhonghua Xueyexue Zazhi
https://www.readbyqxmd.com/read/29562303/dating-the-species-network-allopolyploidy-and-repetitive-dna-evolution-in-american-daisies-melampodium-sect-melampodium-asteraceae
#20
Jamie McCann, Tae-Soo Jang, Jiri Macas, Gerald M Schneeweiss, Nicholas J Matzke, Petr Novák, Tod F Stuessy, José L Villaseñor, Hanna Weiss-Schneeweiss
Allopolyploidy has played an important role in the evolution of the flowering plants. Genome mergers are often accompanied by significant and rapid alterations of genome size and structure via chromosomal rearrangements and altered dynamics of tandem and dispersed repetitive DNA families. Recent developments in sequencing technologies and bioinformatic methods allow for a comprehensive investigation of the repetitive component of plant genomes. Interpretation of evolutionary dynamics following allopolyploidization requires both the knowledge of parentage and the age of origin of an allopolyploid...
March 19, 2018: Systematic Biology
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