scholarly journals CRISPR/Cas9 Targeted Mutagenesis for Functional Genetics in Maize

Plants ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 723
Author(s):  
Charles T. Hunter
Keyword(s):  
Targeted Mutagenesis ◽  
Loss Of Function ◽  
Efficient Manner ◽  
Functional Genetics ◽  
Homozygous Mutant ◽  
Cost Efficient ◽  
Indispensable Tool ◽  
Simple Assembly ◽  
Simultaneous Delivery ◽  
Genetic Strategies

The CRISPR/Cas9-based system for targeted mutagenesis has become an indispensable tool for functional genetics in plants. CRISPR/Cas9 allows users to generate loss-of-function alleles in genes of interest with precision and in a simple-to-use system. This manuscript outlines important points to consider for experimental design and utilization of CRISPR/Cas9 in targeted mutagenesis in maize. It also introduces the pRGEB32-BAR vector modified for use in maize that allows simultaneous delivery of multiple gRNAs using a simple assembly. Vector selection, gRNA design, genetic strategies, and genotyping approaches are discussed, with an emphasis on achieving isolation of homozygous mutant plants in a time- and cost-efficient manner.

scholarly journals First genome edited poinsettias: targeted mutagenesis of flavonoid 3′-hydroxylase using CRISPR/Cas9 results in a colour shift

2021 ◽  
Author(s):  
Daria Nitarska ◽  
Robert Boehm ◽  
Thomas Debener ◽  
Rares Calin Lucaciu ◽  
Heidi Halbwirth
Keyword(s):  
Transgenic Plants ◽  
Genome Editing ◽  
Ornamental Plants ◽  
Targeted Mutagenesis ◽  
Cloning And Expression ◽  
Euphorbia Pulcherrima ◽  
Loss Of Function ◽  
Wild Type ◽  
Promising Tool ◽  
Reddish Orange

AbstractThe CRISPR/Cas9 system is a remarkably promising tool for targeted gene mutagenesis, and becoming ever more popular for modification of ornamental plants. In this study we performed the knockout of flavonoid 3′-hydroxylase (F3′H) with application of CRISPR/Cas9 in the red flowering poinsettia (Euphorbia pulcherrima) cultivar ‘Christmas Eve’, in order to obtain plants with orange bract colour, which accumulate prevalently pelargonidin. F3′H is an enzyme that is necessary for formation of cyanidin type anthocyanins, which are responsible for the red colour of poinsettia bracts. Even though F3′H was not completely inactivated, the bract colour of transgenic plants changed from vivid red (RHS 45B) to vivid reddish orange (RHS 33A), and cyanidin levels decreased significantly compared with the wild type. In the genetically modified plants, an increased ratio of pelargonidin to cyanidin was observed. By cloning and expression of mutated proteins, the lack of F3′H activity was confirmed. This confirms that a loss of function mutation in the poinsettia F3′H gene is sufficient for obtaining poinsettia with orange bract colour. This is the first report of successful use of CRISPR/Cas9 for genome editing in poinsettia.

Investigation on the Machining of Thick Diamond Films by EDM Together with Mechanical Polishing

2007 ◽  
Vol 24-25 ◽  
pp. 377-382
Author(s):  
Rong Fa Chen ◽  
Dun Wen Zuo ◽  
Yu Li Sun ◽  
Wen Zhuang Lu ◽  
D.S. Li ◽  
...  
Keyword(s):  
Diamond Films ◽  
Polycrystalline Diamond ◽  
Mechanical Polishing ◽  
Polishing Process ◽  
Two Stage ◽  
Efficient Manner ◽  
Raman Scattering Spectroscopy ◽  
Electro Discharge Machining ◽  
Diamond Polishing ◽  
Cost Efficient

Although research on various diamond polishing techniques has been carried for years, some issues still need to be examined in order to facilitate application on large areas in a cost-efficient manner. A compositive technique for machining efficiently thick diamond films prepared by DC plasma arc jet is reported in the present paper. A two-stage polishing was applied on thick polycrystalline diamond films, by employing first electro-discharge machining (EDM) for rough polishing and subsequently mechanical polishing for finishing operations. Experimental results obtained clearly indicate the applicability of the proposed two-stage technique for fabricating transparent diamond films that can be used for the production of X-ray windows. Appropriate etching with EDM is an effective pretreatment method for enhancing the efficiency of rough polishing process in mechanical polishing of thick diamond film. The machined surfaces of diamond films are studied by Scanning Electron Microscope (SEM) and Raman Scattering Spectroscopy (Raman).

Achieving Indonesian Oil and Gas production to 1 million barrels per day by 2030 using Nanoflooding as Novel EOR Method: Dream vs. Reality

2021 ◽  
Author(s):  
L. Hendraningrat
Keyword(s):  
Oil Recovery ◽  
Cost Efficiency ◽  
Oil And Gas ◽  
Gas Production ◽  
Oil Price ◽  
Efficient Manner ◽  
Oil And Gas Production ◽  
Cost Efficient ◽  
Nano Size ◽  
Production Target

In low oil price environments, conducting affordable enhanced oil recovery (EOR) projects can be very challenging. One item of interest for successful future EOR should be in how produced fluids are treated and how to achieve cost-efficiency. Nanoflooding, is an emerging EOR technique, which has attracted deployment in recent years. Meanwhile, Indonesia continues to progress towards the national oil and gas production target of one million barrels per day by 2030. This paper presents the observation of opportunities and challenges of using nanoflooding to enable oil and gas production in Indonesia to achieve its desired targets. The study began by mapping the pain points in major oilfields in Indonesia. We observed and discussed the advantage and limitation of traditional mature EOR techniques, status, and ongoing application of EOR in Indonesia. Then, we briefly explained the main reasons why nanoflooding can be considered for future implementation in accelerating oil production in Indonesia, including a discussion about a successful pilot test. As an emerging EOR technique, nanoflooding can be considered as a cost-efficient technique. Silica-based nanofluid can be accessed in a cost-efficient manner and can be executed from an implementation standpoint considering surface facilities. The mechanism that is introduced can help to displace incremental oil more effectively since it can go inside pore throats due to the nano-size. We observed several recognized benefits and challenges to deploy nanoflooding in Indonesia. Based on this study, nanoflooding is very attractive and has potential to be implemented.

scholarly journals Ultrabithorax Is a Micromanager of Hindwing Identity in Butterflies and Moths

2021 ◽  
Vol 9 ◽  
Author(s):  
Amruta Tendolkar ◽  
Aaron F. Pomerantz ◽  
Christa Heryanto ◽  
Paul D. Shirk ◽  
Nipam H. Patel ◽  
...  
Keyword(s):  
Regulatory Networks ◽  
Targeted Mutagenesis ◽  
Junonia Coenia ◽  
Wing Venation ◽  
Loss Of Function ◽  
Color Patterns ◽  
Wide Range ◽  
Range Of Functions ◽  
Gene Regulatory ◽  
Pyralid Moth

The forewings and hindwings of butterflies and moths (Lepidoptera) are differentiated from each other, with segment-specific morphologies and color patterns that mediate a wide range of functions in flight, signaling, and protection. The Hox gene Ultrabithorax (Ubx) is a master selector gene that differentiates metathoracic from mesothoracic identities across winged insects, and previous work has shown this role extends to at least some of the color patterns from the butterfly hindwing. Here we used CRISPR targeted mutagenesis to generate Ubx loss-of-function somatic mutations in two nymphalid butterflies (Junonia coenia, Vanessa cardui) and a pyralid moth (Plodia interpunctella). The resulting mosaic clones yielded hindwing-to-forewing transformations, showing Ubx is necessary for specifying many aspects of hindwing-specific identities, including scale morphologies, color patterns, and wing venation and structure. These homeotic phenotypes showed cell-autonomous, sharp transitions between mutant and non-mutant scales, except for clones that encroached into the border ocelli (eyespots) and resulted in composite and non-autonomous effects on eyespot ring determination. In the pyralid moth, homeotic clones converted the folding and depigmented hindwing into rigid and pigmented composites, affected the wing-coupling frenulum, and induced ectopic scent-scales in male androconia. These data confirm Ubx is a master selector of lepidopteran hindwing identity and suggest it acts on many gene regulatory networks involved in wing development and patterning.

scholarly journals Rethinking the Role of Lawyers for Children : Child Representation in Canadian Family Relationship Cases

2019 ◽  
Vol 59 (4) ◽  
pp. 787-829
Author(s):  
Nicholas Bala ◽  
Rachel Birnbaum
Keyword(s):  
Family Relationship ◽  
Older Children ◽  
Efficient Manner ◽  
Child Report ◽  
Significant Evidence ◽  
The Family ◽  
Justice Process ◽  
Cost Efficient ◽  
Comprehensive Programs

This article examines various methods for involving children in family and child welfare proceedings, surveys varying approaches in different Canadian jurisdictions to the appointment of counsel for children in these cases, and explores the controversies about the role of counsel for children. While child representation is becoming common, in most provinces it is usually limited to welfare proceedings involving older children. All jurisdictions, however, have some provisions to allow for child participation in family relationship cases. Governments and law societies in Canada should develop more coherent and comprehensive programs and policies to ensure that the views of children are considered in the family justice process in a child-focused and cost-efficient manner. There needs to be more recognition of the role of lawyers in facilitating settlement. Appointment of counsel is, however, not always the best way to involve children ; if the child is willing to share views, in some cases this may be better done by a judicial interview, a Views of the Child Report or an assessment. There should be a presumption that counsel will be an instructional advocate, but if a child lacks the capacity or willingness to instruct counsel, counsel should be an advocate for the rights and interests of the child. Regardless of the role adopted, counsel for a child also has responsibility for introducing significant evidence not otherwise before the court.

scholarly journals Effect of randomly distributed polypropylene fiber reinforcement on the shear behavior of sandy soil

2019 ◽  
Vol 41 (3) ◽  
pp. 151-159
Author(s):  
Mehdi Missoum Benziane ◽  
Noureddine Della ◽  
Sidali Denine ◽  
Sedat Sert ◽  
Said Nouri
Keyword(s):  
Sandy Soil ◽  
Mechanical Characteristics ◽  
Polypropylene Fiber ◽  
Fiber Content ◽  
Polypropylene Fibers ◽  
Normal Stresses ◽  
Efficient Manner ◽  
Cost Efficient ◽  
Shear Box ◽  
Loose Medium

AbstractThe inclusions of geosynthetic materials (fibers, geomembranes and geotextiles) is a new improvement technique that ensures uniformity in the soil during construction. The use of tension resisting discreet inclusions like polypropylene fibers has attracted a significant amount of attention these past years in the improvement of soil performance in a cost-efficient manner. A series of direct shear box tests were conducted on unreinforced and reinforced Chlef sand with different contents of fibers (0, 0.25, 0.5 and0.75%) in order to study the mechanical behavior of sand reinforced with polypropylene fibers. Samples were prepared at three different relative densities 30%, 50% and 80% representing loose, medium dense and dense states,respectively, and performed at normal stresses of 50, 100 and 200 kPa. The experimental results show that the mechanical characteristics are improved with the addition of polypropylene fibers. The inclusion of randomly distributed fibers has a significant effect on the shear strength and dilation of sandy soil. The increase in strength is a function of fiber content, where it has been shown that the mechanical characteristics improve with the increase in fiber content up to 0.75%, this improvement is more significant at a higher normal stress and relative density.

Starting from Year One: The Politics of Health in Nicaragua

1980 ◽  
Vol 10 (4) ◽  
pp. 647-656 ◽  
Author(s):  
José Carlos Escudero
Keyword(s):  
Public Health ◽  
Vital Statistics ◽  
Medical System ◽  
Efficient Manner ◽  
Popular Participation ◽  
Mortality And Morbidity ◽  
Sandinista Revolution ◽  
Cost Efficient ◽  
Health Situation ◽  
And Control

The triumphant Sandinista revolution inherited a health situation characterized by high mortality, low life expectancy, widespread malnutrition, and a medical system limited in scope. The extent of these problems can only be estimated as a result of the somocista government's failure to develop an accurate system of vital statistics. While there are many options available for rapidly decreasing the high levels of mortality and morbidity in Nicaragua, the revolutionary government has chosen a strategy which fuses public health and politics. A health network based on popular participation and control is being formed which should not only decrease the high rates of malnutrition and infectious diseases in a cost-efficient manner, but should increase the strength of the revolution as well.

The Serrate locus of Drosophila and its role in morphogenesis of the wing imaginal discs: control of cell proliferation

Development ◽  
1994 ◽  
Vol 120 (3) ◽  
pp. 535-544 ◽  
Author(s):  
S.A. Speicher ◽  
U. Thomas ◽  
U. Hinz ◽  
E. Knust
Keyword(s):  
Cell Proliferation ◽  
Transmembrane Protein ◽  
Ectopic Expression ◽  
Imaginal Discs ◽  
Specific Cell ◽  
Loss Of Function ◽  
Severe Reduction ◽  
Homozygous Mutant ◽  
Wing Imaginal Discs ◽  
Larval Lethality

The Drosophila gene Serrate encodes a transmembrane protein with 14 EGF-like repeats in its extracellular domain. Here we show that loss-of-function mutations in this gene lead to larval lethality. Homozygous mutant larvae fail to differentiate the anterior spiracles, exhibit poorly developed mouth-hooks and show a severe reduction in the size of the wing and haltere primordia, which is not due to cell death. The few homozygous mutant escapers that pupariate develop into pharate adults that almost completely lack wings and halteres. Clonal analysis in the adult epidermis demonstrates a requirement for Serrate during wing and haltere development. Targeted ectopic expression of Serrate in the imaginal discs using the yeast transcriptional activator Gal4 results in regionally restricted induction of cell proliferation, e.g. the ventral tissues in the case of the wings and halteres. The results suggest that the wild-type function of Serrate is required for the control of position-specific cell proliferation during development of meso- and metathoracic dorsal discs, which in turn exerts a direct effect on morphogenesis.

Production of High Amylose and Resistant Starch Rice Through Targeted Mutagenesis of Starch Branching Enzyme Iib by Crispr/cas9

2021 ◽  
Author(s):  
Hsi-Chao Wang ◽  
Yu-Chia Hsu ◽  
Yong-Pei Wu ◽  
Su-Ying Yeh ◽  
Maurice S. B. Ku
Keyword(s):  
Gene Expression ◽  
Enzyme Activity ◽  
Resistant Starch ◽  
Targeted Mutagenesis ◽  
Rice Starch ◽  
Branching Enzyme ◽  
Starch Branching Enzyme ◽  
Homozygous Mutant ◽  
Mutant Lines ◽  
High Amylose

Abstract Rice is the staple food for half of the world’s population. Starch accounts for 80-90% of the total mass of rice seeds, and rice starch is low in resistant starch (RS) with a high glycemic index (GI). RS has gained important since it is beneficial in preventing various diseases. Starch branching enzyme IIb (SBEIIb) plays a key role in the amylopectin synthesis pathway in the endosperm of cereals. Down-regulation of SBEIIb in several important crops has led to high amylose, high RS and low GI starch. In this study, we mutated OsSBEIIb in the japonica rice cultivar TNG82 through CRISPR/Cas9 and investigated the molecular and physicochemical modifications in OsSBEIIb mutant lines, e.g., gene expression, enzyme activity, apparent amylose content (AAC), RS and GI. As expected, the levels of modification in these starch related traits in heterozygous mutant lines were about half as those of homozygous mutant lines. Gene expression and enzyme activity of OsSBEIIb were down-regulated significantly while AAC and RS contents increased progressively from 17.4% and 0.5% in WT, respectively, to as high as 25.0% and 7.5% in heterozygous mutant lines and 36.0% and 12.0% in homozygous mutant lines. Consequently, with increased RS and decreased rate of reducing sugar production, GI progressively decreased in heterozygous and homozygous mutant rice endosperms by 11% and 28%, respectively. Our results demonstrate that it has huge potential for precise and efficient generation of high RS and low GI rice through CRISPR/Cas9 to provide a more suitable source of starch for type II diabetes.

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