Engineering Papers⌕ Search

SEARCH · Engineering Papers

Results for “Multiplex”

Search indexed NASA NTRS and DOE OSTI research on propulsion, heat transfer, battery materials and energy systems. Follow report and document links to the original sources.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 1,261 records · Page 70
All sourcesNASA NTRSDOE OSTI
DOE OSTI2015

AN EFFICIENT, COMPACT, AND VERSATILE FIBER DOUBLE SCRAMBLER FOR HIGH PRECISION RADIAL VELOCITY INSTRUMENTS

We present the design and test results of a compact optical fiber double-scrambler for high-resolution Doppler radial velocity instruments. This device consists of a single optic: a high-index n ∼ 2 ball lens that exchanges the near and far fields between two fibers. When used in conjunction with octagonal fibers, this device yields very high scrambling gains (SGs) and greatly desensitizes the fiber output from any input illumination variations, thereby stabilizing the instrument profile of the spectrograph and improving the Doppler measurement precision. The system is also highly insensitive to input pupil variations, isolating the spectrograph from telescope illumination variations and seeing changes. By selecting the appropriate glass and lens diameter the highest efficiency is achieved when the fibers are practically in contact with the lens surface, greatly simplifying the alignment process when compared to classical double-scrambler systems. This prototype double-scrambler has demonstrated significant performance gains over previous systems, achieving SGs in excess of 10,000 with a throughput of ∼87% using uncoated Polymicro octagonal fibers. Adding a circular fiber to the fiber train further increases the SG to >20,000, limited by laboratory measurement error. While this fiber system is designed for the Habitable-zone Planet Finder spectrograph, it is more generally applicable to other instruments in the visible and near-infrared. Given the simplicity and low cost, this fiber scrambler could also easily be multiplexed for large multi-object instruments.

79 ASTRONOMY AND ASTROPHYSICS↗
DOE OSTI2015

Measurements of the Sunyaev–Zel’Dovich effect in MACS J0647.7+7015 and MACS J1206.2–0847 at high angular resolution with MUSTANG

We present high resolution (9″) imaging of the Sunyaev–Zel’dovich Effect (SZE) toward two massive galaxy clusters, MACS J0647.7+7015 (z = 0.591) and MACS J1206.2–0847 (z = 0.439). We compare these 90 GHz measurements, taken with the Multiplexed Squid/TES Array at Ninety Gigahertz (MUSTANG ) receiver on the Green Bank Telescope, with generalized Navarro–Frenk–White (gNFW) models derived from Bolocam 140 GHz SZE data as well as maps of the thermal gas derived from Chandra X-ray observations. We adopt a serial-fitting approach, in which gNFW models are first fit to the Bolocam data and then compared to the MUSTANG data to determine an overall best-fit model. For MACS J0647.7+7015, we find a gNFW profile with core slope parameter γ = 0.9 fits the MUSTANG image with χ{sub red}{sup 2}=1.005 and probability to exceed (PTE) = 0.34. For MACS J1206.2–0847, we find γ=0.7, χ{sub red}{sup 2}=0.993, and PTE = 0.70. In addition, we find a significant (>3σ) residual SZE feature in MACS J1206.2–0847 coincident with a group of galaxies identified in Very Large Telescope data and filamentary structure found in a weak-lensing mass reconstruction. We suggest the detected sub-structure may be the SZE decrement from a low mass foreground group or an infalling group. Giant Metrewave Radio Telescope measurements at 610 MHz reveal diffuse extended radio emission to the west, which we posit is either an active galactic nucleus-driven radio lobe, a bubble expanding away from disturbed gas associated with the SZE signal, or a bubble detached and perhaps re-accelerated by sloshing within the cluster. Using the spectroscopic redshifts available, we find evidence for a foreground (z = 0.423) or infalling group, coincident with the residual SZE feature.

79 ASTRONOMY AND ASTROPHYSICS↗
DOE OSTI2015

THE FMOS-COSMOS SURVEY OF STAR-FORMING GALAXIES AT z ∼ 1.6. III. SURVEY DESIGN, PERFORMANCE, AND SAMPLE CHARACTERISTICS

We present a spectroscopic survey of galaxies in the COSMOS field using the Fiber Multi-object Spectrograph (FMOS), a near-infrared instrument on the Subaru Telescope. Our survey is specifically designed to detect the Hα emission line that falls within the H-band (1.6–1.8 μm) spectroscopic window from star-forming galaxies with 1.4 < z < 1.7 and M{sub stellar} ≳ 10{sup 10} M{sub ⊙}. With the high multiplex capability of FMOS, it is now feasible to construct samples of over 1000 galaxies having spectroscopic redshifts at epochs that were previously challenging. The high-resolution mode (R ∼ 2600) effectively separates Hα and [N ii]λ6585, thus enabling studies of the gas-phase metallicity and photoionization state of the interstellar medium. The primary aim of our program is to establish how star formation depends on stellar mass and environment, both recognized as drivers of galaxy evolution at lower redshifts. In addition to the main galaxy sample, our target selection places priority on those detected in the far-infrared by Herschel/PACS to assess the level of obscured star formation and investigate, in detail, outliers from the star formation rate (SFR)—stellar mass relation. Galaxies with Hα detections are followed up with FMOS observations at shorter wavelengths using the J-long (1.11–1.35 μm) grating to detect Hβ and [O iii]λ5008 which provides an assessment of the extinction required to measure SFRs not hampered by dust, and an indication of embedded active galactic nuclei. With 460 redshifts measured from 1153 spectra, we assess the performance of the instrument with respect to achieving our goals, discuss inherent biases in the sample, and detail the emission-line properties. Our higher-level data products, including catalogs and spectra, are available to the community.

79 ASTRONOMY AND ASTROPHYSICS↗
DOE OSTI2024

Layer-by-layer assembly of a [Fe-(pyrazine){Pd(CN) 4 }] spin crossover thin film

Abstract [Fe-(pyrazine){Pd(CN) 4 }] (pyrazine = pz) thin films were fabricated using a layer-by-layer assembly approach, a method known to be tunable, versatile, and scalable, since thin films are better-suited for industrial applications. In this study, [Fe-(pz){Pd(CN) 4 }] powder was synthesized, and the results obtained from a vibrating sample magnetometer verified the presence of an abrupt hysteresis loop with widths of 45 K centered around 300 K, indicating good cooperativity. Super conducting quantum interference device magnetometry results indicated a slow spin transition with temperature but with evidence of hysteresis for thin film samples. X-ray absorption analysis provided further support of the spin crossover behavior but differs from the magnetometry because the spin state transition at the surface differs from the bulk of the thin film. X-ray photoelectron spectroscopy provided some insight into issues with the film deposition process and multiplex fitting was used to further support the claim that the surface of the film is different than the bulk of the film.

McElveen, Kayleigh A. (ORCID:0000000210302913)↗
DOE OSTI2020

Preliminary design overview and performance assessment of the ITER low-field side reflectometer

The design of the ITER low-field side reflectometer (LFSR) has matured to a complete end-to-end preliminary design. LFSR will supply three key plasma measurements: (1) electron density profile, (2) electron density fluctuations, and (3) poloidal rotation. Simultaneous measurements of the three quantities are enabled by an array of six monostatic antennas which inject from an equatorial port on the outboard side of the ITER vessel. Low-loss transmission lines, consisting of corrugated, overmoded waveguide and miter bends, transmit the 30–165 GHz, O- and X-mode signals to and from the ITER plasma. Integrated transmission-line components serve a range of purposes, such as protection from high-power stray radiofrequency radiation, accommodation of transmission-line displacement, and simultaneous measurement of reference and plasma phases during the discharge. Broadband transmission signals are realized by full-band microwave transceivers combined with quasi-optical multiplexing. A field-programmable gate array (FPGA) processor demodulates the profile reflectometer signals, enabling real-time density profile measurements for plasma control system feedback. A full-scale transmission line test facility provides an integrated environment to assess the performance of critical LFSR components. Theoretical modeling together with insertion loss measurements provide the basis for a comprehensive power budget, which accounts for transmitted output power, transmission-line losses, antenna coupling, and plasma effects. Results indicate that high signal-to-noise ratios are achievable with the current design. A synthetic reflectometer model, using real design parameters and baseline ITER profiles, has been developed to estimate the return signal. With evolving microwave and data acquisition technologies, full-band, ultrafast sweeps (<1 μs) will be realizable for ITER.

70 PLASMA PHYSICS AND FUSION TECHNOLOGY↗
DOE OSTI2018

LArPix: demonstration of low-power 3D pixelated charge readout for liquid argon time projection chambers

In this work, we report the demonstration of a low-power pixelated readout system designed for three-dimensional ionization charge detection and digital readout of liquid argon time projection chambers (LArTPCs) . Unambiguous 3D charge readout was achieved using a custom-designed system-on-a-chip ASIC (LArPix) to uniquely instrument each pad in a pixelated array of charge-collection pads. The LArPix ASIC, manufactured in 180 nm bulk CMOS, provides 32 channels of charge-sensitive amplification with self-triggered digitization and multiplexed readout at temperatures from 80 K to 300 K . Using an 832-channel LArPix-based readout system with 3 mm spacing between pads, we demonstrated low-noise (0<50 e - RMS equivalent noise charge) and very low-power (<100 μW/channel) ionization signal detection and readout . The readout was used to successfully measure the three-dimensional ionization distributions of cosmic rays passing through a LArTPC, free from the ambiguities of existing projective techniques. The system design relies on standard printed circuit board manufacturing techniques, enabling scalable and low-cost production of large-area readout systems using common commercial facilities. This demonstration overcomes a critical technical obstacle for operation of LArTPCs in high-occupancy environments, such as the near detector site of the Deep Underground Neutrino Experiment (DUNE) .

47 OTHER INSTRUMENTATION↗
DOE OSTI2020

Design and performance of an in-vacuum, magnetic field mapping system for the Muon g-2 experiment

The Muon g - 2 experiment at Fermilab (E989) aims to measure the anomalous magnetic moment, a μ , of the muon with a precision of 140 parts-per-billion. This requires a precise measurement of both the anomalous spin precession frequency, ω α , of muons stored in a magnetic field of 1.45 T, and a precise measurement of that magnetic field in terms of the shielded proton Larmor frequency, ω' ρ . The measurement of ω' ρ with a total systematic uncertainty of 70 parts-perbillion involves a combination of various nuclear magnetic resonance (NMR) probes. There are 378 probes mounted in fixed locations that constantly monitor field drifts. A water-based, cylindrical calibration probe provides the calibration in terms of the shielded proton Larmor frequency. A crucial element for the multi-step measurement of ω' ρ is the regular mapping of the magnetic field over the muon storage region. The former experiment at Brookhaven National Laboratory (BNL) employed an in-vacuum field mapping system equipped with 17 NMR probes, which was developed by the University of Heidelberg. We have refurbished and upgraded this system with new probes and electronics. The upgrades include the addition of 16-bit, 1 MSPS digitization of the NMR signals, which replaced the hardware-implemented zero-crossing counting of the system at Brookhaven. The digitized signals offer new capabilities in the NMR frequency analysis and its related systematic uncertainties. To sustain the higher data rates, a new communication scheme with time-division multiplexing was implemented to separate the important NMR reference clock from the data communication in order to reach the specifications for the accuracy and stability of the reference clock. A new barcode reader provides more precise azimuthal position determination during the measurement and calibration. While the mechanical systems that move the field mapper inside the storage ring have been mostly refurbished from BNL, the motion control system was completely replaced with a custom-built electronics centered around a commercial Galil motion controller. Both the field mapping NMR system and its motion control were successfully commissioned at Fermilab and have been in reliable operation during the first three data taking periods of the experiment at Fermilab. Finally, this article will provide the details of the upgrades of the field mapper and its performance.

47 OTHER INSTRUMENTATION↗
DOE OSTI2022

Prototype hardware design and testing of the global common module for the global trigger subsystem of the ATLAS phase-II upgrade

We report the High-Luminosity Large Hadron Collider (HL-LHC) will deliver more than ten times the integrated luminosity of the previous runs combined. Meeting its stricter throughput requirements poses new challenges to the Trigger and Data Acquisition (TDAQ) systems of the LHC experiments. Introduced in the framework of the ATLAS experiment’s HL upgrade, the Global Trigger (GT) is a new subsystem which will perform offline-like algorithms on full-granularity calorimeter data. The implementation of the GT’s functionality is firmware-focused and is composed of three layers: multiplexing (or data aggregating), global event processing, and demultiplexing interface to the central trigger processor. Each layer will be composed of several, similar nodes, hosted on replicas of identical hardware, the Global Common Module (GCM), an ATCA front board which is designed to be adopted throughout the entire GT subsystem. This article proceeds from the TWEPP 2021 conference and presents the GCM hardware design, performed in 2020, and focuses on some key results of its extensive testing performed in 2021.

46 INSTRUMENTATION RELATED TO NUCLEAR SCIENCE AND ↗
DOE OSTI2023

Radiation tolerance studies of the HV-mux GaNFETs for the HL-LHC ATLAS ITk Strip detector

For the High-Luminosity upgrade of the LHC, the current ATLAS inner detector will be replaced with a new silicon charged-particle tracker, the ITk, which consists of the ITk Pixel and the ITk Strip subdetector. The high voltage multiplexing (HV-Mux) GaNFETs are radiation-tolerant transistors that permit switching off high voltage to malfunctioning sensors on the ITk Strip modules. To ensure the reliability of the GaNFETs in the high radiation environment expected at the HL-LHC, a sample of the production batch was exposed to gamma radiation. Finally, the GaNFETs were characterized pre-irradiation and post-irradiation, and monitored during irradiation.

46 INSTRUMENTATION RELATED TO NUCLEAR SCIENCE AND ↗
DOE OSTI2024

Development of a strip-shaped X-ray mapping system for 9-cell superconducting cavities

Electrons emitted via field emission during superconducting (SC) radio-frequency (RF) cavity tests at vertical test stands often collide with the iris region inside the cavity, generating X-rays at these locations. In 1.3 GHz 9-cell SC RF cavities designed for the International Linear Collider (ILC), stiffener rings located outside the iris region between cells can interfere with X-ray detection, complicating the precise identification of field emission sites. Hence, in this study, we developed a high-density strip X-ray mapping systems (sX-map) that can be inserted into the iris region of ILC-type 9-cell SC RF cavities. This sX-map facilitates efficient and accurate detection of X-rays generated near the irises, unaffected by the presence of stiffener rings. The sX-map consisted of 32 sensors per strip, with sensors spaced approximately 10 mm apart. It was deployed in every iris of the 9-cell cavity, using a total of 320 sensors. A multiplexer was employed to facilitate the readout of a large number of detectors using a minimal number of signal lines, connecting the strips inside within the vertical test cryostat. In a vertical test conducted at Jefferson Lab (JLab), we demonstrated the capability of sX-map to detect X-rays despite the presence of a stiffener ring. This paper presents the detailed design of the sX-map and the results from the vertical test at JLab.

46 INSTRUMENTATION RELATED TO NUCLEAR SCIENCE AND ↗
DOE OSTI2021

Single-shot complete spatiotemporal measurement of terawatt laser pulses

In this work, we demonstrate, for the first time, a single-shot, complete spatiotemporal measurement of pulses from a terawatt-scale, multi-stage-amplified, low repetition-rate laser source. The ultrashort pulse electric field, E(x,y,z,t), is spatiotemporally complex due to distortions that accrue from multiple chirped-pulse amplifiers, which requires a complete characterization. Meanwhile, the instability of the laser source introduces field profiles that vary significantly from pulse to pulse, which, together with the low repetition-rate (15 shots/hour), requires the use of a single-shot measurement technique. To accomplish the measurements, we used a wavelength-multiplexed, digital-holographic technique called Spatially and Temporally Resolved Intensity and Phase Evaluation Device: Full Information from a Single Hologram, specially tailored to measure picosecond pulses at a wavelength of about 1 μm. Specifically, individual pulses from the compact multipulse terawatt laser were measured, with up to 0.3 J per shot of energy and ~2 ps pulse durations, at 1052 nm. With these measurements, we characterized several major spatiotemporal distortions that affect the peak intensity at the laser focus, as well as the pulse-shape instability on a shot-to-shot basis. Our technique allows detailed diagnosis of laser pulses (especially high-order spatiotemporal distortions) and provides straightforward four-dimensional animations of pulse propagation to a focus.

70 PLASMA PHYSICS AND FUSION TECHNOLOGY↗
DOE OSTI2024

Biosensors for the detection of chorismate and cis,cis -muconic acid in Corynebacterium glutamicum

Abstract Corynebacterium glutamicum ATCC 13032 is a promising microbial chassis for industrial production of valuable compounds, including aromatic amino acids derived from the shikimate pathway. In this work, we developed two whole-cell, transcription factor based fluorescent biosensors to track cis,cis-muconic acid (ccMA) and chorismate in C. glutamicum. Chorismate is a key intermediate in the shikimate pathway from which value-added chemicals can be produced, and a shunt from the shikimate pathway can divert carbon to ccMA, a high value chemical. We transferred a ccMA-inducible transcription factor, CatM, from Acinetobacter baylyi ADP1 into C. glutamicum and screened a promoter library to isolate variants with high sensitivity and dynamic range to ccMA by providing benzoate, which is converted to ccMA intracellularly. The biosensor also detected exogenously supplied ccMA, suggesting the presence of a putative ccMA transporter in C. glutamicum, though the external ccMA concentration threshold to elicit a response was 100-fold higher than the concentration of benzoate required to do so through intracellular ccMA production. We then developed a chorismate biosensor, in which a chorismate inducible promoter regulated by natively expressed QsuR was optimized to exhibit a dose-dependent response to exogenously supplemented quinate (a chorismate precursor). A chorismate–pyruvate lyase encoding gene, ubiC, was introduced into C. glutamicum to lower the intracellular chorismate pool, which resulted in loss of dose dependence to quinate. Further, a knockout strain that blocked the conversion of quinate to chorismate also resulted in absence of dose dependence to quinate, validating that the chorismate biosensor is specific to intracellular chorismate pool. The ccMA and chorismate biosensors were dually inserted into C. glutamicum to simultaneously detect intracellularly produced chorismate and ccMA. Biosensors, such as those developed in this study, can be applied in C. glutamicum for multiplex sensing to expedite pathway design and optimization through metabolic engineering in this promising chassis organism. One-Sentence Summary High-throughput screening of promoter libraries in Corynebacterium glutamicum to establish transcription factor based biosensors for key metabolic intermediates in shikimate and β-ketoadipate pathways.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2020

SPIDERS: overview of the X-ray galaxy cluster follow-up and the final spectroscopic data release

ABSTRACT SPIDERS (The SPectroscopic IDentification of eROSITA Sources) is a large spectroscopic programme for X-ray selected galaxy clusters as part of the Sloan Digital Sky Survey-IV (SDSS-IV). We describe the final data set in the context of SDSS Data Release 16 (DR16): the survey overall characteristics, final targeting strategies, achieved completeness, and spectral quality, with special emphasis on its use as a galaxy cluster sample for cosmology applications. SPIDERS now consists of about 27 000 new optical spectra of galaxies selected within 4000 photometric red sequences, each associated with an X-ray source. The excellent spectrograph efficiency and a robust analysis pipeline yield a spectroscopic redshift measurement success rate exceeding 98 per cent, with a median velocity accuracy of 20 km s−1 (at z = 0.2). Using the catalogue of 2740 X-ray galaxy clusters confirmed with DR16 spectroscopy, we reveal the 3D map of the galaxy cluster distribution in the observable Universe up to z ∼ 0.6. We highlight the homogeneity of the member galaxy spectra among distinct regions of the galaxy cluster phase space. Aided by accurate spectroscopic redshifts and by a model of the sample selection effects, we compute the galaxy cluster X-ray luminosity function and we present its lack of evolution up to z = 0.6. Finally we discuss the prospects of forthcoming large multiplexed spectroscopic programmes dedicated to follow up the next generation of all-sky X-ray source catalogues.

Clerc, N.↗
DOE OSTI2022

Observational window effects on multi-object reverberation mapping

ABSTRACT Contemporary reverberation mapping campaigns are employing wide-area photometric data and high-multiplex spectroscopy to efficiently monitor hundreds of active galactic nuclei (AGNs). However, the interaction of the window function(s) imposed by the observation cadence with the reverberation lag and AGN variability time-scales (intrinsic to each source over a range of luminosities) impact our ability to recover these fundamental physical properties. Time dilation effects due to the sample source redshift distribution introduce added complexity. We present comprehensive analysis of the implications of observational cadence, seasonal gaps, and campaign baseline duration (i.e. the survey window function) for reverberation lag recovery. We find that the presence of a significant seasonal gap dominates the efficacy of any given campaign strategy for lag recovery across the parameter space, particularly for those sources with observed-frame lags above 100 d. Using the Australian Dark Energy Survey as a baseline, we consider the implications of this analysis for the 4MOST/Time-Domain Extragalactic Survey campaign providing concurrent follow-up of the Legacy Survey of Space and Time deep-drilling fields, as well as upcoming programmes. We conclude that the success of such surveys will be critically limited by the seasonal visibility of some potential field choices, but show significant improvement from extending the baseline. Optimizing the sample selection to fit the window function will improve survey efficacy.

79 ASTRONOMY AND ASTROPHYSICS↗
DOE OSTI2020

Programmable gene regulation for metabolic engineering using decoy transcription factor binding sites

Abstract Transcription factor decoy binding sites are short DNA sequences that can titrate a transcription factor away from its natural binding site, therefore regulating gene expression. In this study, we harness synthetic transcription factor decoy systems to regulate gene expression for metabolic pathways in Escherichia coli. We show that transcription factor decoys can effectively regulate expression of native and heterologous genes. Tunability of the decoy can be engineered via changes in copy number or modifications to the DNA decoy site sequence. Using arginine biosynthesis as a showcase, we observed a 16-fold increase in arginine production when we introduced the decoy system to steer metabolic flux towards increased arginine biosynthesis, with negligible growth differences compared to the wild type strain. The decoy-based production strain retains high genetic integrity; in contrast to a gene knock-out approach where mutations were common, we detected no mutations in the production system using the decoy-based strain. We further show that transcription factor decoys are amenable to multiplexed library screening by demonstrating enhanced tolerance to pinene with a combinatorial decoy library. Our study shows that transcription factor decoy binding sites are a powerful and compact tool for metabolic engineering.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2019

De novo design of programmable inducible promoters

Abstract Ligand-responsive allosteric transcription factors (aTF) play a vital role in genetic circuits and high-throughput screening because they transduce biochemical signals into gene expression changes. Programmable control of gene expression from aTF-regulated promoter is important because different downstream effector genes function optimally at different expression levels. However, tuning gene expression of native promoters is difficult due to complex layers of homeostatic regulation encoded within them. We engineered synthetic promoters de novo by embedding operator sites with varying affinities and radically reshaped binding preferences within a minimal, constitutive Escherichia coli promoter. Multiplexed cell-based screening of promoters for three TetR-like aTFs generated with this approach gave rich diversity of gene expression levels, dynamic ranges and ligand sensitivities and were 50- to 100-fold more active over their respective native promoters. Machine learning on our dataset revealed that relative position of the core motif and bases flanking the core motif play an important role in modulating induction response. Our generalized approach yields customizable and programmable aTF-regulated promoters for engineering cellular pathways and enables the discovery of new small molecule biosensors.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2024

Quantitative proteomics reveals extensive lysine ubiquitination and transcription factor stability states in Arabidopsis

Abstract Protein activity, abundance, and stability can be regulated by post-translational modification including ubiquitination. Ubiquitination is conserved among eukaryotes and plays a central role in modulating cellular function; yet, we lack comprehensive catalogs of proteins that are modified by ubiquitin in plants. In this study, we describe an antibody-based approach to enrich ubiquitinated peptides coupled with isobaric labeling to enable quantification of up to 18-multiplexed samples. This approach identified 17,940 ubiquitinated lysine sites arising from 6,453 proteins from Arabidopsis (Arabidopsis thaliana) primary roots, seedlings, and rosette leaves. Gene ontology analysis indicated that ubiquitinated proteins are associated with numerous biological processes including hormone signaling, plant defense, protein homeostasis, and metabolism. We determined ubiquitinated lysine residues that directly regulate the stability of three transcription factors, CRYPTOCHROME-INTERACTING BASIC-HELIX-LOOP-HELIX 1 (CIB1), CIB1 LIKE PROTEIN 2 (CIL2), and SENSITIVE TO PROTON RHIZOTOXICITY1 (STOP1) using in vivo degradation assays. Furthermore, codon mutation of CIB1 to create a K166R conversion to prevent ubiquitination, via CRISPR/Cas9-derived adenosine base editing, led to an early flowering phenotype and increased expression of FLOWERING LOCUS T (FT). These comprehensive site-level ubiquitinome profiles provide a wealth of data for future functional studies related to modulation of biological processes mediated by this post-translational modification in plants.

Biochemistry & Molecular Biology↗
DOE OSTI2024

Heritable, multinucleotide deletions in plants using viral delivery of a repair exonuclease and guide RNAs

Abstract CRISPR/Cas9-mediated mutagenesis typically results in short insertion/deletion mutations, which are often too small to disrupt the function of cis-acting regulatory elements. Here, we describe a highly efficient in planta gene editing approach called VirTREX2-HLDel that achieves heritable multinucleotide deletions in both protein-coding genes and noncoding DNA regulatory elements. VirTREX2-HLDel uses RNA viruses to deliver both the 3 prime repair exonuclease 2 (TREX2) and single-guide RNAs. Our method enables recovery of multiplexed heritable deletions and increases the heritable gene editing frequency at poorly edited sites. We identified functional conservation and divergence of MICRORNA164 (miR164) in Nicotiana benthamiana and tomato (Solanum lycopersicum) using VirTREX2-HLDel and observed previously uncharacterized phenotypes in plants with large deletions at this locus. Our viral delivery method reduces the need for tissue culture and will accelerate the understanding of protein-coding and regulatory regions in plants.

59 BASIC BIOLOGICAL SCIENCES↗