CRISPR/Cas systems confer immunity against invading nucleic acids and phages in bacteria and archaea. CRISPR/Cas13a (known previously as C2c2) is a class 2 type VI-A ribonuclease capable of targeting ...and cleaving single-stranded RNA (ssRNA) molecules of the phage genome. Here, we employ CRISPR/Cas13a to engineer interference with an RNA virus, Turnip Mosaic Virus (TuMV), in plants.
CRISPR/Cas13a produces interference against green fluorescent protein (GFP)-expressing TuMV in transient assays and stable overexpression lines of Nicotiana benthamiana. CRISPR RNA (crRNAs) targeting the HC-Pro and GFP sequences exhibit better interference than those targeting other regions such as coat protein (CP) sequence. Cas13a can also process pre-crRNAs into functional crRNAs.
Our data indicate that CRISPR/Cas13a can be used for engineering interference against RNA viruses, providing a potential novel mechanism for RNA-guided immunity against RNA viruses and for other RNA manipulations in plants.
4,16‐para‐substituted 2.2paracylophanes with naphthalene (PCP‐NAP), anthracene (PCP‐ANTH), and tetraphenylethylene (PCP‐TPE), as new through‐space conjugated dimers, were prepared by the ...Suzuki‐Miyaura cross‐coupling reaction of 4,16‐diboryl2.2paracyclophane and the corresponding bromo derivative using Pd(PPh3)4 as a catalyst and KOH as a base. The synthesized compounds were fully characterized by NMR and HR‐MS, and their photophysical and electrochemical properties were studied. The photoluminescence quantum yield of PCP‐NAP, PCP‐ANTH were determined to be 0.21, 0.50, and for PCP‐TPE in aggregated state 0.31, respectively. PCP‐TPE exhibited aggregation‐induced emission characteristics when the water fraction was greater than 50 % in THF/water mixtures. PCP‐ANTH and PCP‐TPE were also characterized by X‐ray crystallography, PCP‐ANTH crystallizes in centrosymmetric monoclinic space group C2/c and PCP‐TPE crystallizes in the centrosymmetric triclinic space group P‐1 with one molecule residing at the inversion center. The observed properties of these π‐stacked dimers were compared with respect to through‐space conjugation and conjugation length in the structure.
Insights into noncovalent interactions by elucidating the profound impact of through‐space conjugation on the optical properties of 2.2paracyclophane based through‐space conjugated dimers. Through a experimental investigations the research sheds light on the intricate interplay of extended and space conjugation. Moreover, it explores the application of Suzuki‐Miyaura coupling reactions, in tailoring the optical behavior of functional materials.
Patients presenting with herpes zoster (HZ) to emergency departments (EDs) across the United States represent a significant number of visits and have pain that is difficult to manage, sometimes even ...requiring opioid medications for adequate analgesia. Ultrasound-guided nerve blocks (UGNBs) are becoming more integrated into the ED physician's tool box for a multimodal approach to analgesia in various indications. Here we describe a novel use of the transgluteal sciatic UGNB for treatment of HZ pain along the S1 dermatome. A 48-year-old woman presented to the ED with right-sided leg pain associated with a HZ rash. After initially failing non-opioid pain management, the ED physician performed a transgluteal sciatic UGNB for our patient, leading to successful complete resolution of her pain, with no adverse effects reported. Our case highlights the potential role of using the transgluteal sciatic UGNB for analgesia related to HZ-related pain, as well as its potential opioid-sparing benefits. Although UGNBs require a baseline understanding of ultrasound technique for procedural guidance, this skillset has recently been incorporated as core competency within emergency medicine training in the United States. UGNBs should therefore be considered in the multimodal analgesic armamentarium for the ED treatment of HZ pain.
Clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated (Cas) systems are key immune mechanisms helping prokaryotic species fend off RNA and DNA viruses. CRISPR/Cas9 ...has broad applications in basic research and biotechnology and has been widely used across eukaryotic species for genome engineering and functional analysis of genes. The recently developed CRISPR/Cas13 systems target RNA rather than DNA and thus offer new potential for transcriptome engineering and combatting RNA viruses. Here, we used CRISPR/LshCas13a to stably engineer
for interference against the RNA genome of Turnip mosaic virus (TuMV). Our data demonstrate that CRISPR RNAs (crRNAs) guiding Cas13a to the sequences encoding helper component proteinase silencing suppressor (HC-Pro) or GFP target 2 (GFP-T2) provide better interference compared to crRNAs targeting other regions of the TuMV RNA genome. This work demonstrates the exciting potential of CRISPR/Cas13 to be used as an antiviral strategy to obstruct RNA viruses, and encourages the search for more robust and effective Cas13 variants or CRISPR systems that can target RNA.
Summary
Targeted genomic regulation is a powerful approach to accelerate trait discovery and development in agricultural biotechnology. Bacteria and archaea use clustered regularly interspaced short ...palindromic repeats (CRISPRs) and CRISPR‐associated (Cas) regulatory systems for adaptive molecular immunity against foreign nucleic acids introduced by invading phages and conjugative plasmids. The type II CRISPR/Cas system has been adapted for genome editing in many cell types and organisms. A recent study used the catalytically inactive Cas9 (dCas9) protein combined with guide‐RNAs (gRNAs) as a DNA‐targeting platform to modulate gene expression in bacterial, yeast, and human cells. Here, we modified this DNA‐targeting platform for targeted transcriptional regulation in planta by developing chimeric dCas9‐based transcriptional activators and repressors. To generate transcriptional activators, we fused the dCas9 C‐terminus with the activation domains of EDLL and TAL effectors. To generate a transcriptional repressor, we fused the dCas9 C‐terminus with the SRDX repression domain. Our data demonstrate that dCas9 fusion with the EDLL activation domain (dCas9:EDLL) and the TAL activation domain (dCas9:TAD), guided by gRNAs complementary to selected promoter elements, induce strong transcriptional activation on Bs3::uidA targets in plant cells. Further, the dCas9:SRDX‐mediated transcriptional repression of an endogenous gene. Thus, our results suggest that the synthetic transcriptional repressor (dCas9:SRDX) and activators (dCas9:EDLL and dCas9:TAD) can be used as endogenous transcription factors to repress or activate transcription of an endogenous genomic target. Our data indicate that the CRISPR/dCas9 DNA‐targeting platform can be used in plants as a functional genomics tool and for biotechnological applications.
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•FMO, MEP and NBO analyses of N and C-nitroso diphenylamine (DPA)•Enhanced NLO-phoric effect in case C-nitroso over N-nitroso DPA.•Substituent effect on static and dynamic NLO ...properties at para position.•Order of substituent effect on NLO properties: Me < OMe < OEt < NH2 < NMe2.•Highest moleculer (µ.β0 = 570 × 10-30 Debye.esu) and first order hyperpolarizability (β0 = 57.46 × 10-30 esu) achieved in case of CNO-pNMe2-DPA.
The nonlinear optical property of N and C-nitroso diphenylamine is the focus of the current work but the D-π-A skeleton-based para substituted nitroso benzene served as the starting point for this investigation. For p-methoxy nitroso benzene, a maximum β0 of 10.47 x 10-30 esu was attained. Diphenylamine analogue was letter on chosen for further improvement. For these nitroso diphenylamines, substituent effects of electron donating groups (EDGs) at the para position have been studied for the nonlinear optical property (NLO). The NMe2 derivative of C-nitroso was found to have the highest value of hyperpolarizability (β0) of 57.46 × 10-30 esu, including 570 × 10-30 Debye.esu as molecular hyperpolarizability (µ×β0), out of these two nitroso forms with substituents Me, OMe, OEt, NH2, and NMe2 at para position. H < Me < OMe < OEt < NH2 < NMe2 is the ascending effect for the β0 in the N-nitroso whereas H < Me < OMe < NH2 < OEt < NMe2 in the C-nitroso. These β0 value shows 155.29, 73.69, 13.12 fold increment as compared to diphenylamine (DPA), urea and p-nitroaniline (PNA) respectively. Upon increasing the λmax, value of calculated βTDω also increases calculated in TD-DFT calculations (βTD1064 <βTD1340 <βTD1460 < βTD1907). These values have shown a deviation range of 7–38 % in case of N-nitroso and 11–52 % in case of C-nitroso as compared to static values. Whereas βHRSω increases with increase in the optical frequency (βHRS1064 >βHRS1340 >βHRS1460 > βHRS1907). These values have shown a deviation range of 4–91 % in case of N-nitroso and 1to > 100 in case of C-nitroso as compared to static values.
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•Relative charge distribution of TPE-EWGs/EDGs.•Substituent effect on static (β0, µβ0) and dynamic (βTDω and βHRSω) NLO properties.•More pronounced effect of the EWGs as compared to ...EDGs.•Highest β0 (52.73 × 10-30 esu) and µβ0 (288.43 × 10-30 Debye.esu) values in case of TPE-NO followed by TPE-NO2.•67.60 fold enhancement of β0 for TPE-NO as compared to prototype urea.
In this study, tetraphenylethylene (TPE) derivatives with various substituents (F, OH, OMe, CN, NO2, and NO) at the para position were selected for the non-linear optical properties. Non-centrosymmetric behavior and charge distribution of these derivatives were evaluated using natural bond orbital (27.24 kcal/mol orbital interaction energy in case of πC6-C10 → π*N46-O47 for TPE-NO), HOMO-LUMO diagrams, and molecular electrostatic potential analyses (OH, 34.15 kcal/mol and CN, −35.39 kcal/mol). TPE derivatives with electron-withdrawing groups displayed higher hyperpolarizability values (molecular, time and frequency dependent values) compared to electron-donating ones. TPE-NO have shown highest values for static (52.73 × 10-30 esu) and molecular (288.43 × 10-30 Debye.esu). TPE with withdrawing groups showed higher limit of first order hyperpolarizability ranging from 1050 × 10-30 to 1530 × 10-30 esu. βHRS1064 >βHRS1340 >βHRS1460 > βHRS1907 and βTD1064 <βTD1340 <βTD1460 < βTD1907 are the order of computed Hyper-Rayleigh scattering and time-dependent first order dynamic hyperpolarizability respectively.
The estimation of the Direction of Arrival (DoA) is one of the most critical parameters for target recognition, identification and classification. MUltiple SIgnal Classification (MUSIC) is a powerful ...technique for DoA estimation. The algorithm requires complex mathematical operations like the computation of the covariance matrix for the input signals, eigenvalue decomposition and signal peak search. All these signal processing operations make real-time and resource-efficient implementation of the MUSIC algorithm on Field Programmable Gate Arrays (FPGAs) a challenge. In this paper, a novel design approach is proposed for the FPGA-implementation of the MUSIC algorithm. This approach enables a significant reduction in both FPGA resources and latency. In more detail, the proposed design enables the estimation of DoA in real-time scenarios in <inline-formula> <tex-math notation="LaTeX">2\mu </tex-math></inline-formula>sec with 30% to 50% fewer resources as compared to existing techniques.
One important factor for successful disease management is the ability to rapidly and accurately identify the causal agent. Plant viruses cause severe economic losses and pose a serious threat to ...sustainable agriculture. Therefore, optimization of the speed, sensitivity, feasibility, portability, and accuracy of virus detection is urgently needed. Here, we developed a clustered regularly interspaced short palindromic repeats (CRISPR)-based nucleic acid diagnostic method utilizing the CRISPR-Cas12a system for detecting two geminiviruses, tomato yellow leaf curl virus (TYLCV) and tomato leaf curl New Delhi virus (ToLCNDV), which have single-stranded DNA genomes. Our assay detected TYLCV and ToLCNDV in infected plants with high sensitivity and specificity. Our newly developed assay can be performed in ~1 h and provides easy-to-interpret visual readouts using a simple, low-cost fluorescence visualizer, making it suitable for point-of-use applications.
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