scholarly journals Asymmetrical cleavages ofSleeping Beautytransposons generate multiple excised transposon fragments during transposition

2019 ◽  
Author(s):  
Qilin Gu ◽  
Xiaojie Yang ◽  
Qing Li ◽  
Yong Long ◽  
Guili Song ◽  
...  
Keyword(s):  
Sleeping Beauty ◽  
Delivery Vehicle ◽  
Essential Requirement ◽  
The Standard Model ◽  
Integration Sites ◽  
Unequal Rates ◽  
Flanking Sequences ◽  
Cleavage Step ◽  
Selective Integration ◽  
Selection Of

ABSTRACTAlthough theSleeping Beauty(SB) transposon is the most validated DNA transposon used as a gene delivery vehicle in vertebrates, many details of the excision and integration steps in the transposition process are unclear. We have probed in detail the products of the excision step and apparent selective integration of a subset of those products during transposition. The standard model of SB transposase-mediated transposition includes symmetrical cleavages at both ends of the transposon for excision and re-integration in another DNA sequence. In our analysis of excised transposon fragments (ETFs), we found evidence for the requirement of certain flanking sequences for efficient cleavage and a significant rate of asymmetrical cleavage during the excision process that generates multiple ETFs. Our results suggest that the cleavage step by SB transposase is not as precise as indicated in most models. Repair of the donor ends can produce eight footprint sequences (TACTGTA, TACAGTA, TACATA, TACGTA, TATGTA, TACTA, TAGTA and TATA). Our data also suggest that mismatch repair (MMR) is not an essential requirement for footprint formation. Among the twenty liberated ETFs, only eight appear to effectively re-integrate into TA sites distributed across the genome, supporting earlier findings of unequal rates of excision and reintegration during SB transposition. These findings may be important in considerations of efficiency of SB transposon remobilization, selection of TA integration sites and detection of SB excision and integration loci, all of which may be important in human gene therapy.

scholarly journals RNA-guided Retargeting of Sleeping Beauty Transposition in Human Cells

2019 ◽  
Author(s):  
Adrian Kovač ◽  
Csaba Miskey ◽  
Michael Menzel ◽  
Esther Grueso ◽  
Andreas Gogol-Döring ◽  
...  
Keyword(s):  
Genome Engineering ◽  
Sleeping Beauty ◽  
Specific Gene ◽  
Guide Rna ◽  
Gene Insertion ◽  
Genome Wide ◽  
Asymmetric Pattern ◽  
Integration Sites ◽  
Genomic Regions ◽  
Selection Of

ABSTRACTTwo different approaches of genomic modification are currently used for genome engineering and gene therapy: integrating vectors, which can efficiently integrate large transgenes but are unspecific with respect to their integration sites, and nuclease-based approaches, which are highly specific but not efficient at integrating large genetic cargoes. Here we demonstrate biased genome-wide integration of the Sleeping Beauty (SB) transposon by combining it with components of the CRISPR/Cas9 system. We provide proof-of-concept that it is possible to influence the target site selection of SB by fusing it to a catalytically inactive Cas9 (dCas9) and by providing a single guide RNA (sgRNA) against the human Alu retrotransposon. Enrichment of transposon integrations was dependent on the sgRNA, occurred in a relatively narrow, ∼200 bp window around the targeted sites and displayed an asymmetric pattern with a bias towards sites that are downstream of the sgRNA targets. Our data indicate that the targeting mechanism specified by CRISPR/Cas9 forces integration into genomic regions that are otherwise poor targets for SB transposition. Future modifications of this technology may allow the development of methods for efficient and specific gene insertion for precision genetic engineering.

scholarly journals Indirect $$ \mathcal{CP} $$ probes of the Higgs-top-quark interaction: current LHC constraints and future opportunities

2020 ◽  
Vol 2020 (11) ◽  
Author(s):  
Henning Bahl ◽  
Philip Bechtle ◽  
Sven Heinemeyer ◽  
Judith Katzy ◽  
Tobias Klingl ◽  
...  
Keyword(s):  
Higgs Boson ◽  
Standard Model ◽  
Top Quark ◽  
Decay Channel ◽  
New Physics ◽  
Current Data ◽  
Quark Interaction ◽  
The Standard Model ◽  
Vector Bosons ◽  
Selection Of

Abstract The $$ \mathcal{CP} $$ CP structure of the Higgs boson in its coupling to the particles of the Standard Model is amongst the most important Higgs boson properties which have not yet been constrained with high precision. In this study, all relevant inclusive and differential Higgs boson measurements from the ATLAS and CMS experiments are used to constrain the $$ \mathcal{CP} $$ CP -nature of the top-Yukawa interaction. The model dependence of the constraints is studied by successively allowing for new physics contributions to the couplings of the Higgs boson to massive vector bosons, to photons, and to gluons. In the most general case, we find that the current data still permits a significant $$ \mathcal{CP} $$ CP -odd component in the top-Yukawa coupling. Furthermore, we explore the prospects to further constrain the $$ \mathcal{CP} $$ CP properties of this coupling with future LHC data by determining tH production rates independently from possible accompanying variations of the $$ t\overline{t}H $$ t t ¯ H rate. This is achieved via a careful selection of discriminating observables. At the HL-LHC, we find that evidence for tH production at the Standard Model rate can be achieved in the Higgs to diphoton decay channel alone.

A novel T-DNA vector design for selection of transgenic lines with simple transgene integration and stable transgene expression

2005 ◽  
Vol 32 (8) ◽  
pp. 671 ◽  
Author(s):  
Song Chen ◽  
Christopher A. Helliwell ◽  
Li-Min Wu ◽  
Elizabeth S. Dennis ◽  
Narayana M. Upadhyaya ◽  
...  
Keyword(s):  
Transgene Expression ◽  
Marker Gene ◽  
Direct Repeat ◽  
Transgene Silencing ◽  
Selective Marker ◽  
Hairpin Rna ◽  
Transgenic Lines ◽  
Flanking Sequences ◽  
Dna Vector ◽  
Selection Of

Plants transformed with Agrobacterium frequently contain T-DNA concatamers with direct-repeat (d / r) or inverted-repeat (i / r) transgene integrations, and these repetitive T-DNA insertions are often associated with transgene silencing. To facilitate the selection of transgenic lines with simple T-DNA insertions, we constructed a binary vector (pSIV) based on the principle of hairpin RNA (hpRNA)-induced gene silencing. The vector is designed so that any transformed cells that contain more than one insertion per locus should generate hpRNA against the selective marker gene, leading to its silencing. These cells should, therefore, be sensitive to the selective agent and less likely to regenerate. Results from Arabidopsis and tobacco transformation showed that pSIV gave considerably fewer transgenic lines with repetitive insertions than did a conventional T-DNA vector (pCON). Furthermore, the transgene was more stably expressed in the pSIV plants than in the pCON plants. Rescue of plant DNA flanking sequences from pSIV plants was significantly more frequent than from pCON plants, suggesting that pSIV is potentially useful for T-DNA tagging. Our results revealed a perfect correlation between the presence of tail-to-tail inverted repeats and transgene silencing, supporting the view that read-through hpRNA transcript derived from i / r T-DNA insertions is a primary inducer of transgene silencing in plants.

scholarly journals The integration preference of Sleeping Beauty at non-TA site is related to the transposon end sequences

2019 ◽  
Author(s):  
Yiting Zhou ◽  
Guangwei Ma ◽  
Jiawen Yang ◽  
Yabin Guo
Keyword(s):  
Site Selection ◽  
Genomic Dna ◽  
Consensus Sequence ◽  
Mouse Cell ◽  
Sleeping Beauty ◽  
Consensus Sequences ◽  
Target Site Selection ◽  
Target Sites ◽  
End Sequences ◽  
Selection Of

Abstract Background: Sleeping Beauty (SB) transposon had been thought to strictly integrate into TA dinucleotides. Recently, we found that SB also integrates into non-TA sites at a lower frequency. Here we performed further study on the non-TA integration of SB. Results: 1) SB can integrate into non-TA sites in HEK293T cells as well as in mouse cell lines. 2) Both the hyperactive transposase SB100X and the traditional SB11 catalyze integrations at non-TA sites. 3) The consensus sequence of the non-TA target sites only occur at the opposite side of the sequenced junction between transposon end and the genomic sequences, indicating that the integrations at non-TA sites are mainly aberrant integrations. 4) The consensus sequence of the non-TA target sites is corresponding to the transposon end sequence. When the transposon end sequence is mutated, the consensus sequences changed too. Conclusion: The interaction between the SB transposon end and genomic DNA may be involved in the target site selection of the SB integrations at non-TA sites.

scholarly journals Calibration of Manning’s roughness in non-instrumented rural basins using a distributed hydrological model

DYNA ◽  
2019 ◽  
Vol 86 (210) ◽  
pp. 164-173
Author(s):  
Carlos Andrés Caro Camargo ◽  
Oscar Fabián Pacheco Merchán ◽  
Hans Paul Sánchez Tueros
Keyword(s):  
Hydrological Model ◽  
Numerical Study ◽  
Research Work ◽  
Roughness Coefficient ◽  
Distributed Hydrological Model ◽  
The Standard Model ◽  
Saint Venant Equations ◽  
Volume Method ◽  
Best Fit ◽  
Selection Of

The purpose of this research work was to calibrate the Manning's roughness coefficient in rural non-instrumented basins using a distributed hydrological model. The process consisted of the selection of several basins with vegetal cover of forests and grasslands, and its subsequent experimental numerical study, in which the hydrological response hydrograph of each reference basin was obtained from the HEC-HMS software and the hydrograph to be calibrated was the Iber software, which is a hydrodynamic model based on the two-dimensional Saint Venant equations, solved by the finite volume method. Once the calibration process was carried out, the roughness coefficients with the best fit for each basin were identified with increases in precipitation, identifying the limitations of these values and the standard model used.

Selecting models and setting standards for teachers of English in Hong Kong

2002 ◽  
Vol 12 (1) ◽  
pp. 13-37 ◽  
Author(s):  
David Coniam ◽  
Peter Falvey
Keyword(s):  
Hong Kong ◽  
English Language ◽  
Language Ability ◽  
Minimum Standard ◽  
Second Language Teaching ◽  
Benchmark Assessment ◽  
The Past ◽  
Setting Process ◽  
The Standard Model ◽  
Selection Of

The perception of falling standards in education, and in second language teaching in particular, has been a constant refrain for the past 20 years as changing needs and practices affect economic processes and manpower requirements. Within this context, this article deals with the establishment of language standards (‘benchmarks’) for teachers of English in Hong Kong. The article deals with two separate but linked notions: first, the notion of which model of the English language should be used as the standard model for English language teacher benchmark assessment in Hong Kong; second, the level of language ability that will be decided upon in order to establish the proficiency standards expected of teachers of English. The article first describes the background to the setting of language standards for teachers of English in Hong Kong. It then examines the selection of and justification for the model of English selected as the ‘minimum standard’. The following section considers the level of language ability agreed upon as the standard that teachers of English need to attain. The article concludes with an examination of the extent to which the standards that have been agreed upon match the needs of the major stakeholders in the standard-setting process.

Implementation of the CMS-HIG-18-011 analysis in the MadAnalysis 5 framework (exotic Higgs decays via two pseudoscalars with two muons and two b-jets; 35.9 fb−1)

2020 ◽  
pp. 2141003
Author(s):  
Joon-Bin Lee ◽  
Jehyun Lee
Keyword(s):  
Higgs Boson ◽  
Center Of Mass ◽  
Final State ◽  
Proton Proton ◽  
Proton Collisions ◽  
Center Of Mass Energy ◽  
Cms Experiment ◽  
The Standard Model ◽  
Proton Proton Collisions ◽  
Selection Of

We present the implementation in the MadAnalysis 5 framework of the CMS-HIG-18-011 search for exotic decays of the Standard Model Higgs boson, in which the Higgs boson is assumed to decay into a pair of light pseudoscalar [Formula: see text], that then further decay into a di-muon and di-[Formula: see text]-jet final state. This analysis considers proton-proton collisions at a center-of-mass energy of 13 TeV and data collected by the CMS experiment in 2016, with an integrated luminosity of 35.9 fb[Formula: see text]. We present a selection of recast predictions, obtained with MadAnalysis 5 and Delphes 3, that include a few differential distributions, yields, and efficiencies. We show that they agree at a level of a few percent with public CMS results.

PHOTOGRAPHIC RESOLVING POWER

1946 ◽  
Vol 24a (4) ◽  
pp. 15-40 ◽  
Author(s):  
L. E. Howlett
Keyword(s):  
Aerial Photography ◽  
Focal Plane ◽  
Resolving Power ◽  
Optical Systems ◽  
Maximum Information ◽  
Essential Requirement ◽  
Future Design ◽  
The Future ◽  
Selection Of

The nature of photographic resolving power is discussed and attention called to the widespread misconception of it that exists at the present time. Most of the detailed discussion applies specifically to photographic objectives intended for use in aerial photography but the general approach to their photographic resolving power is applicable to studies of the photographic performance of all types of optical systems. An annulus type of target is proposed as more suitable than line targets. A method is given for the selection of the photographic focal plane when the essential requirement of the photographic use is the acquisition of maximum information. General remarks are made on the proper trend to be followed in the future design of photographic objectives. Results are presented on a study of a number of well known types of photographic objectives used for aerial photography.

Evolution of Indirect Restorations for Fixed Prosthodontics: Planning, Preparation and Cementation

2019 ◽  
Vol 8 (3) ◽  
pp. 48-63
Author(s):  
Mahul Patel
Keyword(s):  
Health Care ◽  
Primary Health Care ◽  
Patient Outcomes ◽  
Comprehensive Planning ◽  
Essential Requirement ◽  
Fixed Prosthodontics ◽  
Primary Health ◽  
Dental Practitioners ◽  
Restorative Materials ◽  
Selection Of

The provision of indirect restorations utilising contemporary materials for single or multiple abutment units in primary health care is an essential requirement for all general dental practitioners to ensure predictable patient outcomes. This paper highlights the important considerations for comprehensive planning and adept treatment execution and delivery that can enhance the patient's dental experience and outcome. A selection of clinical cases highlighting established and novel restorative materials utilising conventional and adhesive clinical techniques is presented and discussed.

PROBING THE STANDARD MODEL WITH DILEPTON DECAYS OF TOP QUARK PAIRS AT CDF

2007 ◽  
Vol 22 (38) ◽  
pp. 2873-2884
Author(s):  
DANIEL WHITESON
Keyword(s):  
Standard Model ◽  
Cross Section ◽  
Top Quark ◽  
Production Cross Section ◽  
Production Cross ◽  
Fundamental Theory ◽  
Final State ◽  
Quark Pairs ◽  
The Standard Model ◽  
Selection Of

The dilepton decays of the top quark are a powerful laboratory for probing the Standard Model and searching for hints of a more fundamental theory. We present a detailed analysis of the production cross section and the kinematic qualities of top quark pair candidate events in [Formula: see text] collisions at [Formula: see text] = 1.96 TeV collected by the CDF detector which include two leptons in the final state, suggesting the decay [Formula: see text]. We describe the selection of candidate events to suppress major backgrounds and present the number of observed events over background. As a test of the top quark hypothesis, the kinematics of the events are analyzed via a measurement of M top with unprecedented precision.

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