scholarly journals Chimeric crRNA improves CRISPR–Cas12a specificity in the N501Y mutation detection of Alpha, Beta, Gamma, and Mu variants of SARS-CoV-2

PLoS ONE ◽  
2021 ◽  
Vol 16 (12) ◽  
pp. e0261778
Author(s):  
Jun Yang ◽  
Nilakshi Barua ◽  
Md. Nannur Rahman ◽  
Norman Lo ◽  
Tsz Fung Tsang ◽  
...  

Many CRISPR/Cas platforms have been established for the detection of SARS-CoV-2. But the detection platform of the variants of SARS-CoV-2 is scarce because its specificity is very challenging to achieve for those with only one or a few nucleotide(s) differences. Here, we report for the first time that chimeric crRNA could be critical in enhancing the specificity of CRISPR-Cas12a detecting of N501Y, which is shared by Alpha, Beta, Gamma, and Mu variants of SARS-CoV-2 without compromising its sensitivity. This strategy could also be applied to detect other SARS-CoV-2 variants that differ only one or a few nucleotide(s) differences.

1994 ◽  
Vol 304 (2) ◽  
pp. 641-647 ◽  
Author(s):  
R Zang ◽  
H J Müller ◽  
K Kielbassa ◽  
F Marks ◽  
M Gschwendt

Various murine tissues were tested, by using a protein kinase C-eta-specific antiserum, for the expression of type eta protein kinase C. Brain was found to be the richest source of a type eta isoenzyme. Native protein kinase C-eta was partially purified from the cytosol of murine brain by chromatography on DEAE-Sepharose, hydroxyapatite and protamine-agarose. This procedure resulted in a separation of protein kinase C-eta from the other phorbol 12-myristate 13-acetate (PMA)-responsive isoenzymes (alpha, beta, gamma, delta, epsilon) and allowed, for the first time, characterization of the native enzyme. The protein kinase C of type eta from mouse brain is a phospholipid-dependent Ca(2+)-unresponsive protein kinase. Both PMA and bryostatin activate the kinase for phosphorylation of a substrate as well as for autophosphorylation. Various pseudo-substrate-related peptides are suitable as substrates for the eta-type kinase, peptide delta being the best and peptides eta and epsilon the poorest substrates. The enzyme is inhibited by staurosporine and staurosporine-related compounds, such as K252a and Gö 6976. However, protein kinase C-eta, like protein kinase C-delta, is around two orders of magnitude less sensitive towards Gö 6976 than are the Ca(2+)-responsive isoenzymes (alpha, beta, gamma). The eta-type protein kinase C exhibits an extreme tendency to lose its PMA-responsiveness. Consequently, purification of the enzyme to homogeneity has not yet been successful.


2016 ◽  
Vol 52 (11) ◽  
pp. 2405-2407 ◽  
Author(s):  
N. Kolevzon ◽  
D. Hashoul ◽  
S. Naik ◽  
A. Rubinstein ◽  
E. Yavin

SNP detection of the mutated kRAS oncogene in living cells is shown to be achieved for the first time by cell-permeable and far-red emitting PNA–FIT probes.


2019 ◽  
Vol 9 (10) ◽  
pp. 2106 ◽  
Author(s):  
Santos Villafaina ◽  
Daniel Collado-Mateo ◽  
Juan P. Fuentes-García ◽  
Francisco J. Domínguez-Muñoz ◽  
Narcís Gusi

Fibromyalgia is a chronic syndrome that is characterized by widespread pain and an altered brain dynamic. The aim of this study was to analyze the effect of the duration of the symptoms on the cortical activity of women with fibromyalgia using electroencephalogram power spectrum analyses in an eye-closed resting state. Twenty-nine women participated in this cross-sectional study (N: 29; age: 55.59 [9.50]). Theta, alpha, beta-1, beta-2, and beta-3 frequency bands were analyzed using EEGLAB. Theta power significantly correlated with the duration of the symptoms, but not with age. In addition, participants were divided into two groups according to number the years for which they were suffering from fibromyalgia. Participants who had a longer duration of symptoms obtained higher theta power in the frontal (Fp1, F4, F7, F8, and Fz), central (C3, C4, and Cz), and parietal (P3 and Pz) areas than those who had a shorter duration of symptoms, which may be related to brain aging. This exploratory study demonstrates for the first time that the frontal, central, and parietal areas may be influenced by the years in which they were suffering from the symptoms of fibromyalgia. This might indicate that the duration of these symptoms may have a higher impact on brain aging than the actual age of the patient.


2021 ◽  
Vol 0 (0) ◽  
Author(s):  
Anthony Tromba

Abstract We prove for the first time that classical Morse theory applies to functionals of the form 𝒥 ⁢ ( u ) = 1 2 ⁢ ∫ Ω A α ⁢ β i ⁢ j ⁢ ( x ) ⁢ ∂ ⁡ u i ∂ ⁡ x α ⁢ ∂ ⁡ u j ∂ ⁡ x β ⁢ 𝑑 x + ∫ Ω G ⁢ ( x , u ) ⁢ 𝑑 x \displaystyle\mathcal{J}(u)=\frac{1}{2}\int_{\Omega}A^{ij}_{\alpha\beta}(x)% \frac{\partial u^{i}}{\partial x^{\alpha}}\frac{\partial u^{j}}{\partial x^{% \beta}}\,dx+\int_{\Omega}G(x,u)\,dx where u : Ω → ℝ N {u:\Omega\to\mathbb{R}^{N}} , Ω ⊂ ℝ n {\Omega\subset\mathbb{R}^{n}} compact with C ∞ {C^{\infty}} boundary ∂ ⁡ Ω {\partial\Omega} , u | ∂ ⁡ Ω = φ {u|_{\partial\Omega}=\varphi} , and we argue that this is the largest class to which Morse theory applies.


Author(s):  
S. Fujishiro

The mechanical properties of three titanium alloys (Ti-7Mo-3Al, Ti-7Mo- 3Cu and Ti-7Mo-3Ta) were evaluated as function of: 1) Solutionizing in the beta field and aging, 2) Thermal Mechanical Processing in the beta field and aging, 3) Solutionizing in the alpha + beta field and aging. The samples were isothermally aged in the temperature range 300° to 700*C for 4 to 24 hours, followed by a water quench. Transmission electron microscopy and X-ray method were used to identify the phase formed. All three alloys solutionized at 1050°C (beta field) transformed to martensitic alpha (alpha prime) upon being water quenched. Despite this heavily strained alpha prime, which is characterized by microtwins the tensile strength of the as-quenched alloys is relatively low and the elongation is as high as 30%.


Author(s):  
J. Chakraborty ◽  
A. P. Sinha Hikim ◽  
J. S. Jhunjhunwala

Although the presence of annulate lamellae was noted in many cell types, including the rat spermatogenic cells, this structure was never reported in the Sertoli cells of any rodent species. The present report is based on a part of our project on the effect of torsion of the spermatic cord to the contralateral testis. This paper describes for the first time, the fine structural details of the annulate lamellae in the Sertoli cells of damaged testis from guinea pigs.One side of the spermatic cord of each of six Hartly strain adult guinea pigs was surgically twisted (540°) under pentobarbital anesthesia (1). Four months after induction of torsion, animals were sacrificed, testes were excised and processed for the light and electron microscopic investigations. In the damaged testis, the majority of seminiferous tubule contained a layer of Sertoli cells with occasional spermatogonia (Fig. 1). Nuclei of these Sertoli cells were highly pleomorphic and contained small chromatinic clumps adjacent to the inner aspect of the nuclear envelope (Fig. 2).


Author(s):  
M. Rühle ◽  
J. Mayer ◽  
J.C.H. Spence ◽  
J. Bihr ◽  
W. Probst ◽  
...  

A new Zeiss TEM with an imaging Omega filter is a fully digitized, side-entry, 120 kV TEM/STEM instrument for materials science. The machine possesses an Omega magnetic imaging energy filter (see Fig. 1) placed between the third and fourth projector lens. Lanio designed the filter and a prototype was built at the Fritz-Haber-Institut in Berlin, Germany. The imaging magnetic filter allows energy-filtered images or diffraction patterns to be recorded without scanning using efficient area detection. The energy dispersion at the exit slit (Fig. 1) results in ∼ 1.5 μm/eV which allows imaging with energy windows of ≤ 10 eV. The smallest probe size of the microscope is 1.6 nm and the Koehler illumination system is used for the first time in a TEM. Serial recording of EELS spectra with a resolution < 1 eV is possible. The digital control allows X,Y,Z coordinates and tilt settings to be stored and later recalled.


Author(s):  
Z.L. Wang ◽  
J. Bentley ◽  
R.E. Clausing ◽  
L. Heatherly ◽  
L.L. Horton

Microstructural studies by transmission electron microscopy (TEM) of diamond films grown by chemical vapor deposition (CVD) usually involve tedious specimen preparation. This process has been avoided with a technique that is described in this paper. For the first time, thick as-grown diamond films have been examined directly in a conventional TEM without thinning. With this technique, the important microstructures near the growth surface have been characterized. An as-grown diamond film was fractured on a plane containing the growth direction. It took about 5 min to prepare a sample. For TEM examination, the film was tilted about 30-45° (see Fig. 1). Microstructures of the diamond grains on the top edge of the growth face can be characterized directly by transmitted electron bright-field (BF) and dark-field (DF) images and diffraction patterns.


Author(s):  
Shou-kong Fan

Transmission and analytical electron microscopic studies of scale microstructures and microscopic marker experiments have been carried out in order to determine the transport mechanism in the oxidation of Ni-Al alloy. According to the classical theory, the oxidation of nickel takes place by transport of Ni cations across the scale forming new oxide at the scale/gas interface. Any markers deposited on the Ni surface are expected to remain at the scale/metal interface after oxidation. This investigation using TEM transverse section techniques and deposited microscopic markers shows a different result,which indicates that a considerable amount of oxygen was transported inward. This is the first time that such fine-scale markers have been coupled with high resolution characterization instruments such as TEM/STEM to provide detailed information about evolution of oxide scale microstructure.


Author(s):  
Yimei Zhu ◽  
J. Tafto

The electron holes confined to the CuO2-plane are the charge carriers in high-temperature superconductors, and thus, the distribution of charge plays a key role in determining their superconducting properties. While it has been known for a long time that in principle, electron diffraction at low angles is very sensitive to charge transfer, we, for the first time, show that under a proper TEM imaging condition, it is possible to directly image charge in crystals with a large unit cell. We apply this new way of studying charge distribution to the technologically important Bi2Sr2Ca1Cu2O8+δ superconductors.Charged particles interact with the electrostatic potential, and thus, for small scattering angles, the incident particle sees a nuclei that is screened by the electron cloud. Hence, the scattering amplitude mainly is determined by the net charge of the ion. Comparing with the high Z neutral Bi atom, we note that the scattering amplitude of the hole or an electron is larger at small scattering angles. This is in stark contrast to the displacements which contribute negligibly to the electron diffraction pattern at small angles because of the short g-vectors.


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