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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.

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At least 19 records
All sourcesNASA NTRSDOE OSTI
DOE OSTI2022

Expanding the Scope of Bacterial CRISPR Activation with PAM-Flexible dCas9 Variants

CRISPR-Cas transcriptional tools have been widely applied for programmable regulation of complex biological networks. In comparison to eukaryotic systems, bacterial CRISPR activation (CRISPRa) has stringent target site requirements for effective gene activation. While genes may not always have an NGG protospacer adjacent motif (PAM) at the appropriate position, PAM-flexible dCas9 variants can expand the range of targetable sites. Here we systematically evaluate a panel of PAM-flexible dCas9 variants for their ability to activate bacterial genes. We observe that dxCas9-NG provides a high dynamic range of gene activation for sites with NGN PAMs while dSpRY permits modest activity across almost any PAM. Similar trends were observed for heterologous and endogenous promoters. For all variants tested, improved PAM-flexibility comes with the trade-off that CRISPRi-mediated gene repression becomes less effective. Weaker CRISPR interference (CRISPRi) gene repression can be partially rescued by expressing multiple sgRNAs to target many sites in the gene of interest. Furthermore, our work provides a framework to choose the most effective dCas9 variant for a given set of gene targets, which will further expand the utility of CRISPRa/i gene regulation in bacterial systems.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2022

Spacer2PAM: A computational framework to guide experimental determination of functional CRISPR-Cas system PAM sequences

Abstract RNA-guided nucleases from CRISPR-Cas systems expand opportunities for precise, targeted genome modification. Endogenous CRISPR-Cas systems in many prokaryotes are attractive to circumvent expression, functionality, and unintended activity hurdles posed by heterologous CRISPR-Cas effectors. However, each CRISPR-Cas system recognizes a unique set of protospacer adjacent motifs (PAMs), which requires identification by extensive screening of randomized DNA libraries. This challenge hinders development of endogenous CRISPR-Cas systems, especially those based on multi-protein effectors and in organisms that are slow-growing or have transformation idiosyncrasies. To address this challenge, we present Spacer2PAM, an easy-to-use, easy-to-interpret R package built to predict and guide experimental determination of functional PAM sequences for any CRISPR-Cas system given its corresponding CRISPR array as input. Spacer2PAM can be used in a ‘Quick’ method to generate a single PAM prediction or in a ‘Comprehensive’ method to inform targeted PAM libraries small enough to screen in difficult to transform organisms. We demonstrate Spacer2PAM by predicting PAM sequences for industrially relevant organisms and experimentally identifying seven PAM sequences that mediate interference from the Spacer2PAM-informed PAM library for the type I-B CRISPR-Cas system from Clostridium autoethanogenum. We anticipate that Spacer2PAM will facilitate the use of endogenous CRISPR-Cas systems for industrial biotechnology and synthetic biology.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2020

Programming PAM antennae for efficient CRISPR-Cas9 DNA editing

Bacterial CRISPR-Cas9 nucleases have been repurposed as powerful genome editing tools. Whereas engineering guide RNAs or Cas nucleases have proven to improve the efficiency of CRISPR editing, modulation of protospacer-adjacent motif (PAM), indispensable for CRISPR, has been less explored. Here, we develop a DNA origami–based platform to program a PAM antenna microenvironment and address its performance at the single-molecule level with submolecular resolution. To mimic spatially controlled in vivo PAM distribution as may occur in chromatin, we investigate the effect of PAM antennae surrounding target DNA. We find that PAM antennae effectively sensitize the DNA cleavage by recruiting Cas9 molecules. Super-resolution tracking of single single-guide RNA/Cas9s reveals localized translocation of Cas9 among spatially proximal PAMs. We find that the introduction of the PAM antennae effectively modulates the microenvironment for enhanced target cleavage (up to ~50%). These results provide insight into factors that promote more efficient genome editing.

59 BASIC BIOLOGICAL SCIENCES↗
NASA NTRS2004

Autonomous NanoTechnology Swarm (ANTS) Prospecting Asteroid Mission (PAM), Asteroid Proximity Operations

The Autonomous NanoTechnology Swarm (ANTS) is a generic mission architecture based on spatially distributed spacecraft, autonomous and redundant components, and hierarchical organization. The ANTS Prospecting Asteroid Mission (PAM) is an ANTS application which will nominally use a swarm of 1000 spacecraft. There would be 10 types of "specialists" with common spacecraft buses. There would be 10 subswarms of approximately 100 spacecraft each or approximately 10 of each specialist in each swarm. The ANTS PAM primary objective is the exploration of the asteroid belt in search of resources and material with astrobiologically relevant origins and signatures. The ANTS PAM spacecraft will nominally be released from a station in an Earth-Moon L1 libration point orbit, and they will use Solar sails for propulsion. The sail structure would be highly flexible, capable of changing morphology to change cross-section for capture of sunlight or to form effective "tip vanes" for attitude control. ANTS PAM sails would be capable of full to partial deployment, to change effective sail area and center of pressure, and thus allow attitude control. Results of analysis of a transfer trajectory from Earth to a sample target asteroid will be presented. ANTS PAM will require continuous coverage of different asteroid locations as close as one to two asteroid "diameters" from the surface of the asteroid for periods of science data collection during asteroid proximity operations. Hovering spacecraft could meet the science data collection objectives. The results of hovering analysis will be presented. There are locations for which hovering is not possible, for example on the illuminated side of the asteroid. For cases where hovering is not possible, the results of utilizing asteroid formations to orbit the asteroid and achieve the desired asteroid viewing will be presented for sample asteroids. The ability of ANTS PAM to reduce the area of the solar sail during asteroid proximity operations is critical to the maintenance of orbiting formations for a period of time. Results of analysis of potential "traffic" problems during asteroid proximity operations will be presented.

Marr, Greg↗
DOE OSTI2023

EB Vs PAM Vs VAR for Uranium Alloys

Questions were raised during the February ’97 PDP Meeting at LLNL as to the advisability of funding three separate melting development programs (Advanced Vacuum Arc Remelt (VAR), Plasma Arc Melting (PAM) and Electron Beam Melting (EB)) through PDP in this era of shrinking research funds. The main issues seemed to be: 1) Have we evaluated the potentials of the three processes sufficiently to eliminate any of them out of hand and 2) Have any of these processes to date produced results which show clear advantages over the other two. The feeling at the meeting toward the latter seemed to be that all three processes required further development before they would be accepted by the complex as the standard. As far as evaluating potential for process improvements, however, we at Livermore did go through somewhat of a trade study in 1993 when we proposed the EB route. It seemed to us that even with the elimination of the skull caster via a VIM/VAR/VAR route, the problem of recycle limitations in the VIM step due to excessive carbon pick-up limited the potential for a great improvement in material utilization using this method. A single-step, cold hearth route looked to be the preferable choice. Both EB and PAM are used commercially as cold hearth processes, and both have been used for the production of refractory and specialty metals; EB since the mid 1950’s and PAM since the mid 1980’s. Both seem to have found their individual niches with some applications being suited to EB and others to PAM. Both can be adapted to continuous casting techniques and both are capable of imparting the high heat fluxes necessary to melt refractory metals. The major differences appear to be that for most applications, the EB process is capable of producing a purer product, while PAM results in less loss of volatile components. There exists an abundance of information in the open literature on the results of processing via both routes on various materials. The attached 1984 paper gives a good explanation of electron beam melting and plasma melting, and, I believe, a fair assessment of the advantages and limitations of both processes.

36 MATERIALS SCIENCE↗
DOE OSTI2021

Flexible‐rate optoelectronic wireless transmission system at 101 GHz using multiband CAP‐PAM modulation and envelope detection

Abstract In this paper, we have experimentally demonstrated the first flexible‐rate optoelectronic terahertz wireless transmission system at 101 GHz based on multiband CAP‐PAM modulation and cost‐efficient envelope detection. Instead of traditional M‐QAM mapping, PAM‐N mapping is used in CAP modulation with low‐computational complexity. The system performances and optimal capacity in both single‐band case and multiband case are intensively investigated. The results show that the highest data rate for single‐band system is 13.35 Gbit/s with CAP‐PAM‐5 modulation, while the optimum aggregate transmission line rate of 8.49 Gbit/s can be obtained with 4‐sub‐bands CAP‐PAM‐{5,3,2,2} modulation under 7% FEC limit of 3.8 × 10 −3 for four‐user access.

Zhang, Jiao↗
NASA NTRS1982

Effects of miso- and mesoscale obstructions on PAM winds obtained during project NIMROD

Data from 27 PAM (Portable Automated Mesonet) stations, operational as a phase of project NIMROD (Northern Illinois Meteorological Research on Downburst), are presented. It was found that PAM-measured winds are influenced by the mesoscale obstruction of the Chicago metropolitan area, as well as by the misoscale obstruction of identified trees and buildings. The mesoscale obstruction was estimated within the range of near zero to 50%, increasing toward the city limits, while the misoscale obstruction was estimated as being as large as 58% near obstructing trees which were empirically calculated to cause a wind speed deficit 50-80 times their height. Despite a statistical analysis based on one-million PAM winds, wind speed and stability transmission factors could not be accurately calculated; thus, in order to calculate the airflow free from obstacle, PAM-measured winds must be corrected.

Fujita, T. T.↗
NASA NTRS1985

Ground processing of the McDonnell Douglas Payload Assist Module (PAM)

The payload assist module (PAM) ground processing operations which have evolved since they were started in 1982 are described. The objective of the changes was to reduce the prelaunch testing of the airborne support equipment to increase the throughput of PAM systems while not compromising the reliability of the system when functioned on-orbit. The changes that resulted from the initial cargo element ground processing, the on-orbit performance of the systems, plus the postflight refurbishment and recertification of the airborne support equipment resulted in significant reductions in labor expenditures and work shifts required to prepare a PAM system for flight.

Bryan, C. E.↗
NASA NTRS1982

Software system for reducing PAM-2 data

A software system for reducing PAM-II data was constructed. The data reduction process concatenates data tapes; determines ephemeris; and inverts full sun extinction data. Tests of this data reduction process show that PAM-II data can be compared with data from other, similar satellites.

Pepin, T. J.↗
NASA NTRS1983

Low-level microburst wind structure using Doppler radar and PAM data

The Joint Airport Weather Studies (JAWS) observation network used at an airport in Denver, CO consisted of one S-band and two C-band radars, together with 27 Portable Automated Mesonet (PAM) stations. Each of the three microburst case studies observed during the JAWS experiment occurred within the surface PAM network, and these surface data were compared with the radar analyses. It emerges that if microburst vertical motions are to be accurately resolved, the spacing of the surface stations must be considerably less than 4 km.

Kessinger, C.↗
DOE OSTI2020

PAM module for OAuth 2.0 Device flow

This is a PAM module that lets you log in via SSH to servers using OpenID Connect credentials, instead of SSH Keys or a username and password combination. It uses the OAuth2 Device Flow, which means that during the login process, you will click a link and log in to your OpenID Connect Provider, which will then authenticate you for the SSH session. This module will then check if you're in the right group(s) or have a specified username, and allow or deny access.

Surkont, Jarosław↗
NASA NTRS1983

Microbursts in JAWS depicted by Doppler radars, PAM, and aerial photographs

Preliminary results obtained from the JAWS (Joint Airport Weather Studies) Project near Denver, Colorado in the spring and summer of 1982 using Doppler radar, PAM, and aerial photography are presented. The definitions of the microburst phenomenon are discussed, and statistics comparing NIMROD (Northern Illinois Meteorological Research On Downbursts) for the Midwest region are compared with JAWS for the High Plains region. Possible parent clouds of the microburst are considered, and an analysis of a macroburst/microburst event on July 14, 1982 is presented.

Fujita, T. T.↗
NASA NTRS2003

Exploring with PAM: Prospecting ANTS Missions for Solar System Surveys

ANTS (Autonomous Nano-Technology Swarm), a large (1000 member) swarm of nano to picoclass (10 to 1 kg) totally autonomous spacecraft, are being developed as a NASA advanced mission concept. ANTS, based on a hierarchical insect social order, use an evolvable, self-similar, hierarchical neural system in which individual spacecraft represent the highest level nodes. ANTS uses swarm intelligence attained through collective, cooperative interactions of the nodes at all levels of the system. At the highest levels this can take the form of cooperative, collective behavior among the individual spacecraft in a very large constellation. The ANTS neural architecture is designed for totally autonomous operation of complex systems including spacecraft constellations. The ANTS (Autonomous Nano Technology Swarm) concept has a number of possible applications. A version of ANTS designed for surveying and determining the resource potential of the asteroid belt, called PAM (Prospecting ANTS Mission), is examined here.

Clark, P. E.↗