scholarly journals Gene editing in Brassica napus for basic research and trait development

2021 ◽  
Author(s):  
Greg F. W. Gocal
Keyword(s):  
Brassica Napus ◽  
Gene Editing ◽  
Basic Research ◽  
Oilseed Crop ◽  
Delivery Methods ◽  
Grain Composition ◽  
History Of ◽  
Pod Shatter ◽  
New Alleles ◽  
Trait Development

AbstractThe genome of Brassica napus L. is the result of several polyploidization events that occurred during the history of B. napus. Due to its relatively short domestication history, diversity is relatively limited. An increasing number of loci in this crop’s genome have been gene-edited using various technologies and reagent delivery methods for basic research as well as for trait development. New alleles have been developed as edits in single, 2, 4, or more homologous loci in this important oilseed crop. This comprehensive review will summarize new alleles that have been developed as they relate to weed control, flowering, self-incompatibility, plant hormone biology, disease resistance, grain composition, and pod shatter reduction. These new alleles have significantly augmented our understanding of both plant growth and development for basic research as well as for their potential commercial impacts.

scholarly journals Spring flowering habit in field pennycress (Thlaspi arvense) has arisen multiple independent times

2017 ◽  
Author(s):  
Kevin M. Dorn ◽  
Evan B. Johnson ◽  
Erin Daniels ◽  
Donald L. Wyse ◽  
M. David Marks
Keyword(s):  
Cold Season ◽  
Oilseed Crop ◽  
Loss Of Function ◽  
Thlaspi Arvense ◽  
New Information ◽  
Annual Habit ◽  
History Of ◽  
Winter Annual ◽  
The Difference ◽  
New Alleles

Summary- Pennycress (Thlaspi arvense L.) is currently being developed as a new cold-season oilseed crop. Like many Brassicaceae, pennycress can exhibit either a winter or spring annual phenotype. In Arabidopsis, mutations in negative regulators of flowering, including FLOWERING LOCUS C (FLC) and FRIGIDA can cause the transition to a spring annual habit. The genetics underlying the difference between spring and winter annual pennycress are currently unknown.- Using whole genome sequencing across wild spring annual pennycress accessions, co-segregation analyses, and comparative genomics approaches, we identify new alleles of TaFLC and explore their geographic distribution.- We report that loss of function mutations in TaFLC confer the spring annual phenotype in pennycress. We have also identified four natural alleles of TaFLC that confer a spring annual growth habit. The two spring annual FLC alleles present in European accessions were only identified in accessions collected in Montana, USA.- In pennycress, the spring annual habit has arisen several independent times. Accessions harboring the two European alleles were introduced to North America, likely after the species became a widespread on the continent. These findings provide new information on the natural history of the introduction and spread of the spring annual phenotype in pennycress.

scholarly journals Effective editing for lysophosphatidic acid acyltransferase 2/5 in allotetraploid rapeseed (Brassica napus L.) using CRISPR-Cas9 system

2019 ◽  
Vol 12 (1) ◽  
Author(s):  
Kai Zhang ◽  
Liluo Nie ◽  
Qiqi Cheng ◽  
Yongtai Yin ◽  
Kang Chen ◽  
...  
Keyword(s):  
Brassica Napus ◽  
Lysophosphatidic Acid ◽  
Oil Content ◽  
Gene Editing ◽  
Oil Production ◽  
Oilseed Crop ◽  
Oil Bodies ◽  
Lysophosphatidic Acid Acyltransferase ◽  
Homologous Genes ◽  
Mutant Lines

Abstract Background Brassica napus is one of the most important oilseed crops, and can supply considerable amounts of edible oil as well as provide raw materials for the production of biodiesel in the biotechnology industry. Lysophosphatidic acid acyltransferase (LPAT), a key enzyme in the Kennedy pathway, catalyses fatty acid chains into 3-phosphoglycerate and promotes further production of oil in the form of triacylglycerol. However, because B. napus is an allotetraploid with two subgenomes, the precise genes which involved in oil production remain unclear due to the intractability of efficiently knocking out all copies with high genetic redundancy. Therefore, a robust gene editing technology is necessary for gene function analysis. Results An efficient gene editing technology was developed for the allotetraploid plant B. napus using the CRISPR-Cas9 system. Previous studies showed poor results in either on-target or off-target activity in B. napus. In the present study, four single-gRNAs and two multi-gRNAs were deliberately designed from the conserved coding regions of BnLPAT2 which has seven homologous genes, and BnLPAT5, which has four homologous genes. The mutation frequency was found to range from 17 to 68%, while no mutation was observed in the putative off-target sites. The seeds of the Bnlpat2/Bnlpat5 mutant were wizened and showed enlarged oil bodies, disrupted distribution of protein bodies and increased accumulation of starch in mature seeds. The oil content decreased, with an average decrease of 32% for Bnlpat2 lines and 29% for Bnlpat5 lines in single-gRNA knockout lines, and a decline of 24% for Bnlpat2 mutant lines (i.e., g123) and 39% for Bnlpat2/Bnlpat5 double mutant lines (i.e., g134) in multi-gRNA knockout lines. Conclusions Seven BnLPAT2 homologous genes and four BnLPAT5 homologous genes were cleaved completely using the CRISPR-Cas9 system, which indicated that it is effective for editing all homologous genes in allotetraploid rapeseed, despite the relatively low sequence identities of both gene families. The size of the oil bodies increased significantly while the oil content decreased, confirming that BnLPAT2 and BnLPAT5 play a role in oil biosynthesis. The present study lays a foundation for further oil production improvement in oilseed crop species.

scholarly journals New Insights into the Therapeutic Applications of CRISPR/Cas9 Genome Editing in Breast Cancer

Genes ◽  
2021 ◽  
Vol 12 (5) ◽  
pp. 723
Author(s):  
Munazza Ahmed ◽  
Grace Hope Daoud ◽  
Asmaa Mohamed ◽  
Rania Harati
Keyword(s):  
Breast Cancer ◽  
Dna Sequence ◽  
Gene Editing ◽  
Causes Of Death ◽  
Breast Cancer Treatment ◽  
Delivery Methods ◽  
Therapeutic Applications ◽  
Tumorigenic Potential ◽  
Its Gene ◽  
Shed Light

Breast cancer is one of the most prevalent forms of cancer globally and is among the leading causes of death in women. Its heterogenic nature is a result of the involvement of numerous aberrant genes that contribute to the multi-step pathway of tumorigenesis. Despite the fact that several disease-causing mutations have been identified, therapy is often aimed at alleviating symptoms rather than rectifying the mutation in the DNA sequence. The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 is a groundbreaking tool that is being utilized for the identification and validation of genomic targets bearing tumorigenic potential. CRISPR/Cas9 supersedes its gene-editing predecessors through its unparalleled simplicity, efficiency and affordability. In this review, we provide an overview of the CRISPR/Cas9 mechanism and discuss genes that were edited using this system for the treatment of breast cancer. In addition, we shed light on the delivery methods—both viral and non-viral—that may be used to deliver the system and the barriers associated with each. Overall, the present review provides new insights into the potential therapeutic applications of CRISPR/Cas9 for the advancement of breast cancer treatment.

‘Pod shatter’ in Arabidopsis thaliana Brassica napus and B. juncea

1996 ◽  
Vol 181 (2) ◽  
pp. 195-203 ◽  
Author(s):  
J. SPENCE ◽  
Y. VERCHER ◽  
P. GATES ◽  
N. HARRIS
Keyword(s):  
Arabidopsis Thaliana ◽  
Brassica Napus ◽  
Pod Shatter

scholarly journals Breaking in and busting out: cell-penetrating peptides and the endosomal escape problem

2017 ◽  
Vol 8 (3-4) ◽  
pp. 131-141 ◽  
Author(s):  
Julia C. LeCher ◽  
Scott J. Nowak ◽  
Jonathan L. McMurry
Keyword(s):  
Cell Penetrating Peptides ◽  
Endosomal Escape ◽  
Great Promise ◽  
Delivery Methods ◽  
Oligomeric Proteins ◽  
Cell Penetrating ◽  
History Of ◽  
A Cell ◽  
Escape Problem ◽  
Primary Focus

AbstractCell-penetrating peptides (CPPs) have long held great promise for the manipulation of living cells for therapeutic and research purposes. They allow a wide array of biomolecules from large, oligomeric proteins to nucleic acids and small molecules to rapidly and efficiently traverse cytoplasmic membranes. With few exceptions, if a molecule can be associated with a CPP, it can be delivered into a cell. However, a growing realization in the field is that CPP-cargo fusions largely remain trapped in endosomes and are eventually targeted for degradation or recycling rather than released into the cytoplasm or trafficked to a desired subcellular destination. This ‘endosomal escape problem’ has confounded efforts to develop CPP-based delivery methods for drugs, enzymes, plasmids, etc. This review provides a brief history of CPP research and discusses current issues in the field with a primary focus on the endosomal escape problem, for which several promising potential solutions have been developed. Are we on the verge of developing technologies to deliver therapeutics such as siRNA, CRISPR/Cas complexes and others that are currently failing because of an inability to get into cells, or are we just chasing after another promising but unworkable technology? We make the case for optimism.

A Climate for Science Policy

2015 ◽  
Vol 45 (4) ◽  
pp. 577-609 ◽  
Author(s):  
Matthew L. Wallace
Keyword(s):  
Science Policy ◽  
Scientific Community ◽  
Basic Research ◽  
Internal Conflict ◽  
Atmospheric Sciences ◽  
Federal Organization ◽  
History Of ◽  
New Policies ◽  
Policies And Programs ◽  
Second World

Led by the Meteorological Service of Canada, atmospheric research in Canada underwent a period of rapid growth after the end of the Second World War. Within this federal organization, and in response to operational challenges and staff shortages, there were significant investments in basic research and in research oriented toward external users within Canada. Specifically, new policies and programs were put in place to enable the organization to gain legitimacy within the scientific community and within the federal government. New links with stakeholders and, more importantly, the development of explicit policies to guide research were a prime focus. These formalized strategies for pursuing two parallel types of research generated some internal conflict, but also helped form a common scientific identity among personnel. There were concerted efforts to disseminate research products and reinforce links both with the scientific community and with external users of meteorological and climatological research. Borne out by quantitative data, this science policy–centered history sheds light on the development of research and research specializations in the field in Canada. Most importantly, it provides insight into the global postwar expansion of the atmospheric sciences, which is strongly tied to national contexts. Indeed, the quest for legitimacy and the close connection to government priorities is central to the history of the atmospheric sciences in the twentieth century. More broadly, this case study points to a possible new conception of government science driven by political, bureaucratic, and scientific imperatives, as a means to shed light on scientific networks and practices.

scholarly journals Efficient Protoplast Regeneration Protocol and CRISPR/Cas9-Mediated Editing of Glucosinolate Transporter (GTR) Genes in Rapeseed (Brassica napus L.)

2021 ◽  
Vol 12 ◽  
Author(s):  
Xueyuan Li ◽  
Sjur Sandgrind ◽  
Oliver Moss ◽  
Rui Guan ◽  
Emelie Ivarson ◽  
...  
Keyword(s):  
Brassica Napus ◽  
Shoot Regeneration ◽  
Gene Editing ◽  
Protoplast Culture ◽  
Callus Formation ◽  
Protoplast Regeneration ◽  
Induction Medium ◽  
Shoot Induction ◽  
Brassica Napus L ◽  
High Concentrations

Difficulty in protoplast regeneration is a major obstacle to apply the CRISPR/Cas9 gene editing technique effectively in research and breeding of rapeseed (Brassica napus L.). The present study describes for the first time a rapid and efficient protocol for the isolation, regeneration and transfection of protoplasts of rapeseed cv. Kumily, and its application in gene editing. Protoplasts isolated from leaves of 3–4 weeks old were cultured in MI and MII liquid media for cell wall formation and cell division, followed by subculture on shoot induction medium and shoot regeneration medium for shoot production. Different basal media, types and combinations of plant growth regulators, and protoplast culture duration on each type of media were investigated in relation to protoplast regeneration. The results showed that relatively high concentrations of NAA (0.5 mg l−1) and 2,4-D (0.5 mg l−1) in the MI medium were essential for protoplasts to form cell walls and maintain cell divisions, and thereafter auxin should be reduced for callus formation and shoot induction. For shoot regeneration, relatively high concentrations of cytokinin were required, and among all the combinations tested, 2.2 mg l−1 TDZ in combination with auxin 0.5 mg l−1 NAA gave the best result with up to 45% shoot regeneration. Our results also showed the duration of protoplast culture on different media was critical, as longer culture durations would significantly reduce the shoot regeneration frequency. In addition, we have optimized the transfection protocol for rapeseed. Using this optimized protocol, we have successfully edited the BnGTR genes controlling glucosinolate transport in rapeseed with a high mutation frequency.

scholarly journals International Relations of Medieval Europe in Works of D. I. Kachenovskiy (1827–1872)

2019 ◽  
Keyword(s):  
International Relations ◽  
International Law ◽  
Middle Ages ◽  
Research Work ◽  
Basic Research ◽  
Positive Influence ◽  
Substantial Impact ◽  
Feudal Society ◽  
History Of ◽  
The Middle Ages

The article dwells on reaearch and academic activity of D. I. Kachenovskiy at Kharkiv University. His contribution is shown to the study of international relations history at Middle Ages. It is emphasized that we are quite right to call D. I. Kachenovskiy one of most prominent Ukrainian experts in this field, despite his certain idealization of feudal society. Main teaching subjects of D. I. Kachenovskiy were international law and state law of European powers. The basic research work by D. I. Kachenovskiy became his doctoral thesis «On Privateers and Prize Proceedings Relative to Neutral Merchandise» (1855) which became widely known both in Russian Empire and Europe. D. I. Kachenovskiy admitted the appropriateness of privateering, characterized its essence and shared the opinion of well-known French scholar J.-M. Pardessus that charter by Aragon king Alfonso III of 1288 had been the first legislative act relating to privateering. The first period in European privateering history when “naval war was almost indistinguishable from piracy” lasted, by the scholar’s opinion, up to the end of XVI century. His «Course in International Law» was published in 1863, though unfinished, and embraced the antiquity as well as Middle Ages. The concept of international law development in the Middle Ages as proposed by D. I. Kachenovskiy included several basic elements. Those are: admission of the tremendous contribution of Christian Church, Roman Popes and German Emperors in elaboration of this law, influence of public order and national character of German tribes, as well as statement of general détente of mores and international tension in the Middle Ages under the effect of above-cited factors. The scholar emphasized positive influence of knighthood on customs of war and international relations. Medieval international law, history of privateering and trade, social-political and cultural life of Florence and effect of internal factors on its development were the main lines in research of the scientist. Works by D. I. Kachenovskiy exerted the substantial impact not only on students, but on his colleagues as well, and his scientific treatments mostly sustained time exam.

scholarly journals GENE THERAPY FOR HAEMOPHILIA

2021 ◽  
Vol 2021 (1) ◽  
Author(s):  
A Asghar ◽  
Z Asjad ◽  
H Tahir ◽  
Z Maheen ◽  
S Hanif
Keyword(s):  
Gene Therapy ◽  
Gene Editing ◽  
Genetic Disorders ◽  
Coagulation Factor ◽  
Retroviral Vectors ◽  
Adeno Associated Virus ◽  
General Application ◽  
Blood Disorder ◽  
Statistical Consideration ◽  
History Of

The blood disorder, Hemophilia, has its roots embedded deep into the history of genetic disorders. The European royal family is one of the most prominent families to be affected by this disease thus, dubbing it 'the royal disease'. The types of Hemophilia are divided into two based on the type of coagulation factor mutation found in the patient. For treating haemophilia, gene therapy is done by using different vectors such as lentiviral and retroviral vectors but due to the production of limited expression different adeno associated virus (AAV) strains are used. Some engineerly modified vectors are currently used to get the best possible results. The clinical trials prove the efficacy of these vectors so through their obtained statistical consideration, patient experience and population study once can design vaccines and drugs for haemophilia patients but also due to pre-existing Nabs and pre-existing HCV or HBV infection, the general application of AAV gene therapy is currently limited. The possibility of gene editing for the repair of the mutation is on the horizon.

Sign in / Sign up

Export Citation Format

Share Document