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NASA NTRS2009

Validity of the Aluminum Equivalent Approximation in Space Radiation Shielding

The origin of the aluminum equivalent shield approximation in space radiation analysis can be traced back to its roots in the early years of the NASA space programs (Mercury, Gemini and Apollo) wherein the primary radiobiological concern was the intense sources of ionizing radiation causing short term effects which was thought to jeopardize the safety of the crew and hence the mission. Herein, it is shown that the aluminum equivalent shield approximation, although reasonably well suited for that time period and to the application for which it was developed, is of questionable usefulness to the radiobiological concerns of routine space operations of the 21 st century which will include long stays onboard the International Space Station (ISS) and perhaps the moon. This is especially true for a risk based protection system, as appears imminent for deep space exploration where the long-term effects of Galactic Cosmic Ray (GCR) exposure is of primary concern. The present analysis demonstrates that sufficiently large errors in the interior particle environment of a spacecraft result from the use of the aluminum equivalent approximation, and such approximations should be avoided in future astronaut risk estimates. In this study, the aluminum equivalent approximation is evaluated as a means for estimating the particle environment within a spacecraft structure induced by the GCR radiation field. For comparison, the two extremes of the GCR environment, the 1977 solar minimum and the 2001 solar maximum, are considered. These environments are coupled to the Langley Research Center (LaRC) deterministic ionized particle transport code High charge (Z) and Energy TRaNsport (HZETRN), which propagates the GCR spectra for elements with charges (Z) in the range I <= Z <= 28 (H -- Ni) and secondary neutrons through selected target materials. The coupling of the GCR extremes to HZETRN allows for the examination of the induced environment within the interior' of an idealized spacecraft as approximated by a spherical shell shield, and the effects of the aluminum equivalent approximation for a good polymeric shield material such as genetic polyethylene (PE). The shield thickness is represented by a 25 g/cm spherical shell. Although one could imagine the progression to greater thickness, the current range will be sufficient to evaluate the qualitative usefulness of the aluminum equivalent approximation. Upon establishing the inaccuracies of the aluminum equivalent approximation through numerical simulations of the GCR radiation field attenuation for PE and aluminum equivalent PE spherical shells, we Anther present results for a limited set of commercially available, hydrogen rich, multifunctional polymeric constituents to assess the effect of the aluminum equivalent approximation on their radiation attenuation response as compared to the generic PE.

Badavi, Francis F.↗

NASA NTRSDate not supplied

Robust Trajectory Optimization and GN&C Performance Analysis for NRHO Rendezvous

This paper evaluates several candidate Near-Rectilinear Halo Orbits (NRHO) rendezvous trajectory designs using linear covariance (LinCov) analysis and determines the optimal locations for NRHO rendezvous translational maneuver locations. The performance of several candidate relative trajectory designs are determined as a function of relative navigation accuracy (angles only), inertial optical navigation (OpNav), range observability maneuvers, maneuver execution errors, relative maneuver targeting, and environment uncertainties. Further, the optimal locations of rendezvous maneuvers are determined for each of the candidate reference trajectories. The long-term goal of this research is to utilize LinCov and a genetic optimization algorithm (GA) to determine a complete end-to-end optimal NRHO trajectory design that is robust to navigation errors, maneuver execution errors, and environment uncertainties. This paper represents a first step toward this goal. Three candidate rendezvous trajectories with varying numbers of range-observability maneuvers are evaluated for their robustness to uncertainties, errors, and total trajectory correction delta-v performance. Some key elements of this analysis include relative navigation performance in an NRHO, relative trajectory dispersion performance, and total 3-sigma delta-v performance. This development provides the foundation to then determine an optimal and robust end-to-end NRHO rendezvous trajectory, including the determination of the optimal locations of range observability maneuvers, if needed.

Linear Covariance Analysis↗

NASA NTRS2000

Piezoceramic Actuator Placement for Acoustic Control of Panels

Optimum placement of multiple traditional piezoceramic actuators is determined for active structural acoustic control of flat panels. The structural acoustic response is determined using acoustic radiation filters and structural surface vibration characteristics. Linear Quadratic Regulator (LQR) control is utilized to determine the optimum state feedback gain for active structural acoustic control. The optimum actuator location is determined by minimizing the structural acoustic radiated noise using a modified genetic algorithm. Experimental tests are conducted and compared to analytical results. Anisotropic piezoceramic actuators exhibit enhanced performance when compared to traditional isotropic piezoceramic actuators. As a result of the inherent isotropy, these advanced actuators develop strain along the principal material axis. The orientation of anisotropic actuators is investigated on the effect of structural vibration and acoustic control of curved and flat panels. A fully coupled shallow shell finite element formulation is developed to include anisotropic piezoceramic actuators for shell structures.

Bevan, Jeffrey S.↗

NASA NTRS2001

Piezoceramic Actuator Placement for Acoustic Control of Panels

Optimum placement of multiple traditional piezoceramic actuators is determined for active structural acoustic control of flat panels. The structural acoustic response is determined using acoustic radiation filters and structural surface vibration characteristics. Linear Quadratic Regulator (LQR) control is utilized to determine the optimum state feedback gain for active structural acoustic control. The optimum actuator location is determined by minimizing the structural acoustic radiated noise using a modified genetic algorithm. Experimental tests are conducted and compared to analytical results. Anisotropic piezoceramic actuators exhibits enhanced performance when compared to traditional isotropic piezoceramic actuators. As a result of the inherent isotropy, these advanced actuators develop strain along the principal material axis. The orientation of anisotropic actuators is investigated on the effect of structural vibration and acoustic control of curved and flat panels. A fully coupled shallow shell finite element formulation is developed to include anisotropic piezoceramic actuators for shell structures.

Bevan, Jeffrey S.↗

NASA NTRSDate not supplied

The maize rough endosperm6 (rgh6) mutant encodes a predicted DEAD-box RNA helicase

Maize rough endosperm (rgh) mutants have defective seeds with a rough, etched, or pitted endosperm surface. Molecular genetic analysis of this mutant class has identified multiple RNA processing proteins critical to endosperm development. We isolated the rgh6 locus from the UniformMu transposon tagging population. Mutant kernels have reduced grain-fill with defective embryos that fail to germinate. Self-pollination of rgh6 heterozygotes produces mutant seeds at frequency consistent with a single recessive mutation. Molecular mapping localized rgh6 to a 60 kbp interval on chromosome 5. PCR analysis of gene models within the fine-map interval identified a Mutator (Mu) transposon insertion within a predicted DEAD-box RNA helicase gene. A second allele of rgh6 was identified from UniformMu population. Reciprocal crosses between rgh6-umu1 and rgh6-umu2 heterozygotes produces mutant seeds at a frequency consistent with a 3:1 ratio indicating these mutants are allelic. Both rgh6-umu1 and rgh6-umu2 have Mu element insertions at the same insertion site within the DEAD-box RNA helicase gene. However, the terminal inverted repeat sequences are polymorphic between the alleles showing the alleles are independent mutations. We conclude that mutation of the DEAD-box RNA helicase causes the rgh6 phenotype. Further characterization of the rgh6 mutant phenotype and biochemical investigation of the RGH6 protein is expected to give further insights on the roles of RNA processing in endosperm development.

maize↗

DOE OSTI2024

An interpretable model of pre-mRNA splicing for animal and plant genes

Pre-mRNA splicing is a fundamental step in gene expression, conserved across eukaryotes, in which the spliceosome recognizes motifs at the 3' and 5' splice sites (SSs), excises introns, and ligates exons. SS recognition and pairing is often influenced by protein splicing factors (SFs) that bind to splicing regulatory elements (SREs). Here, we describe SMsplice, a fully interpretable model of pre-mRNA splicing that combines models of core SS motifs, SREs, and exonic and intronic length preferences. We learn models that predict SS locations with 83 to 86% accuracy in fish, insects, and plants and about 70% in mammals. Learned SRE motifs include both known SF binding motifs and unfamiliar motifs, and both motif classes are supported by genetic analyses. Our comparisons across species highlight similarities between non-mammals, increased reliance on intronic SREs in plant splicing, and a greater reliance on SREs in mammalian splicing.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2025

Regulation of sarcomere formation and function in the healthy heart requires a titin intronic enhancer

Heterozygous truncating variants in the sarcomere protein titin (TTN) are the most common genetic cause of heart failure. To understand mechanisms that regulate abundant cardiomyocyte (CM) TTN expression, we characterized highly conserved intron 1 sequences that exhibited dynamic changes in chromatin accessibility during differentiation of human CMs from induced pluripotent stem cells (hiPSC-CMs). Homozygous deletion of these sequences in mice caused embryonic lethality, whereas heterozygous mice showed an allele-specific reduction in Ttn expression. A 296 bp fragment of this element, denoted E1, was sufficient to drive expression of a reporter gene in hiPSC-CMs. Deletion of E1 downregulated TTN expression, impaired sarcomerogenesis, and decreased contractility in hiPSC-CMs. Site-directed mutagenesis of predicted binding sites of NK2 homeobox 5 (NKX2-5) and myocyte enhancer factor 2 (MEF2) within E1 abolished its transcriptional activity. In embryonic mice expressing E1 reporter gene constructs, we validated in vivo cardiac-specific activity of E1 and the requirement for NKX2-5- and MEF2-binding sequences. Moreover, isogenic hiPSC-CMs containing a rare E1 variant in the predicted MEF2-binding motif that was identified in a patient with unexplained dilated cardiomyopathy (DCM) showed reduced TTN expression. Together, these discoveries define an essential, functional enhancer that regulates TTN expression. Manipulation of this element may advance therapeutic strategies to treat DCM caused by TTN haploinsufficiency.

Kim, Yuri↗

DOE OSTI2022

Can fractal dimensions objectivize gastropod shell morphometrics? A case study from Lake Lugu (SW China)

1. Morphometrics are fundamental for the analysis of size and shape in fossils, particularly because soft parts or DNA are rarely preserved and hard parts such as shells are commonly the only source of information. Geometric morphometrics, that is, landmark analysis, is well established for the description of shape but it exhibits a couple of shortcomings resulting from subjective choices during landmarking (number and position of landmarks) and from difficulties in resolving shape at the level of micro-sculpture. 2. With the aid of high-resolution 3D scanning technology and analyses of fractal dimensions, we test whether such shortcomings of linear and landmark morphometrics can be overcome. As a model group, we selected a clade of modern viviparid gastropods from Lake Lugu, with shells that show a high degree of sculptural variation. Linear and landmark analyses were applied to the same shells in order to establish the fractal dimensions. 3. The genetic diversity of the gastropod clade was assessed. The genetic results suggest that the gastropod clade represents a single species. The results of all morphometric methods applied are in line with the genetic results, which is that no specific morphotype could be delimited. Apart from this overall agreement, landmark and fractal dimension analyses do not correspond to each other but represent data sets with different information. Generally, the fractal dimension values quantify the roughness of the shell surface, the resolution of the 3D scans determining the level. In our approach, we captured the micro-sculpture but not the first-order sculptural elements, which explains that fractal dimension and landmark data are not in phase. 4. We can show that analyzing fractal dimensions of gastropod shells opens a window to more detailed information that can be considered in evolutionary and ecological contexts. We propose that using low-resolution 3D scans may successfully substitute landmark analyses because it overcomes the subjective landmarking. Analyses of 3D scans with higher resolution than used in this study will provide surface roughness information at the mineralogical level. We suggest that fractal dimension analyses of a combination of differently resolved 3D models will significantly improve the quality of shell morphometrics.

54 ENVIRONMENTAL SCIENCES↗

DOE OSTI2021

Mechanism of Rad26-assisted rescue of stalled RNA polymerase II in transcription-coupled repair

Transcription-coupled repair is essential for the removal of DNA lesions from the transcribed genome. The pathway is initiated by CSB protein binding to stalled RNA polymerase II. Mutations impairing CSB function cause severe genetic disease. Yet, the ATP-dependent mechanism by which CSB powers RNA polymerase to bypass certain lesions while triggering excision of others is incompletely understood. Here we build structural models of RNA polymerase II bound to the yeast CSB ortholog Rad26 in nucleotide-free and bound states. This enables simulations and graph-theoretical analyses to define partitioning of this complex into dynamic communities and delineate how its structural elements function together to remodel DNA. We identify an allosteric pathway coupling motions of the Rad26 ATPase modules to changes in RNA polymerase and DNA to unveil a structural mechanism for CSB-assisted progression past less bulky lesions. Our models allow functional interpretation of the effects of Cockayne syndrome disease mutations.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2024

Clocking out and letting go to unleash green biotech applications in a photosynthetic host

Cyanobacteria are photosynthetic bacteria whose gene expression patterns are globally regulated by their circadian (daily) clocks. Due to their ability to use sunlight as their energy source, they are also attractive hosts for “green” production of pharmaceuticals, renewable fuels, and chemicals. However, despite the application of traditional genetic tools such as the identification of strong promoters to enhance the expression of heterologous genes, cyanobacteria have lagged behind other microorganisms such as Escherichia coli and yeast as economically efficient cell factories. The previous approaches have ignored large-scale constraints within cyanobacterial metabolic networks on transcription, predominantly the pervasive control of gene expression by the circadian (daily) clock. Here, we show that reprogramming gene expression by releasing circadian repressor elements in the transcriptional regulatory pathways coupled with inactivation of the central oscillating mechanism enables a dramatic enhancement of expression in cyanobacteria of heterologous genes encoding both catalytically active enzymes and polypeptides of biomedical significance.

Science & Technology - Other Topics↗

NASA NTRS1977

KREEP petrogenesis revisited

An experimentally determined crystallization sequence for a composition similar to the LKFM breccias and experimentally determined crystal/liquid distribution coefficients are used to calculate the evolution of the abundances of several key elements during the crystallization of LKFM liquids. Results indicate that the LKFM breccias are not themselves precursors to Apollo 15 igneous KREEP, but that compositions slightly poorer in Ti and Fe might be. Consideration of the evolution of Ti and Sm abundances during the differentiation of proposed whole-moon compositions suggests that it may be necessary to appeal to 'unorthodox' mechanisms such as magma hybridization or interaction of magma with genetically unrelated minerals within the lunar crust to produce KREEP-rich liquids having the observed Ti/Sm. It is concluded that the compositions of samples with Sm contents greater than those in LKFM are dominated by igneous processes, while samples with lower Sm contents may be mixtures of an anorthositic component, a Mg-rich Sm-poor component, and a component similar in composition to LKFM.

Mckay, G. A.↗

DOE OSTI2021

Characterization of Mammalian In Vivo Enhancers Using Mouse Transgenesis and CRISPR Genome Editing [Book Chapter]

Embryonic morphogenesis is strictly dependent on tight spatiotemporal control of developmental gene expression, which is typically achieved through the concerted activity of multiple enhancers driving cell type-specific expression of a target gene. Mammalian genomes are organized in topologically associated domains, providing a preferred environment and framework for interactions between transcriptional enhancers and gene promoters. While epigenomic profiling and three-dimensional chromatin conformation capture have significantly increased the accuracy of identifying enhancers, assessment of subregional enhancer activities via transgenic reporter assays in mice remains the gold standard for assigning enhancer activity in vivo. Once this activity is defined, the ideal method to explore the functional necessity of a transcriptional enhancer and its contribution to target gene dosage and morphological or physiological processes is deletion of the enhancer sequence from the mouse genome. Here we present detailed protocols for efficient introduction of enhancer-reporter transgenes and CRISPR-mediated genomic deletions into the mouse genome, including a step-by-step guide for pronuclear microinjection of fertilized mouse eggs. We provide instructions for the assembly and genomic integration of enhancer-reporter cassettes that have been used for validation of thousands of putative enhancer sequences accessible through the VISTA enhancer browser, including a recently published method for robust site-directed transgenesis at the H11 safe-harbor locus. Together, these methods enable rapid and large-scale assessment of enhancer activities and sequence variants in mice, which is essential to understand mammalian genome function and genetic diseases.

cis-regulatory elements↗

DOE OSTI2022

Hubbard-corrected oxide formation enthalpies without adjustable parameters

A density functional theory (DFT) approach to computing transition metal oxide heat of formation without adjustable parameters is presented. Different degrees of d-electron localization in oxides are treated within the DFT+U approach with site-dependent, first-principles Hubbard U-parameters obtained from linear response theory, and delocalized states in the metallic phases are treated without Hubbard corrections. Comparison of relative stabilities of these differently treated phases is enabled by a local d-electron density matrix-dependent model, which was found by genetic programming against experimental reference formation enthalpies. This mathematically simple model does not explicitly depend on the Hubbard-corrected ionic species and is shown to reproduce the heats of formation of the Mott insulators Ca 2 RuO 4 and Y 2 Ru 2 O 7 within ~3% of experimental results, where the experimental training data did not contain Ru oxides. This newly developed method thus absolves from the need for element-specific corrections fitted to experiments in existing Hubbard-corrected approaches to the prediction of reaction energies of transition metal oxides and metals. The absence of fitting parameters opens up here the possibility to predict relative thermodynamic stabilities and reaction energies involving d-states of varying degree of localization at transition metal oxide interfaces and defects, where site-dependent U-parameters will be particularly important and devising a fitting scheme against experimental data with predictive power would be exceedingly difficult.

transition metal oxides↗

NASA NTRS1996

On the Relationship between the Apollo 16 Ancient Regolith Breccias and Feldspathic Fragmental Breccias, and the Composition of the Prebasin Crust in the Central Highlands of the Moon

Two types of texturally and compositionally similar breccias that consist largely of fragmental debris from meteorite impacts occur at the Apollo 16 lunar site: Feldspathic fragmental breccias (FFBS) and ancient regolith breccias (ARBs). Both types of breccia are composed of a suite of mostly feldspathic components derived from the early crust of the Moon and mafic impact-melt breccias produced during the time of basin formation. The ARBs also contain components, such as agglutinates and glass spherules, indicating that the material of which they are composed occurred at the surface of the Moon as fine-grained regolith prior to lithification of the breccias. These components are absent from the FFBS, suggesting that the FFBs might be the protolith of the ARBS. However, several compositional differences exist between the two types of breccia, making any simple genetic relationship implausible. First, clasts of mafic impact-melt breccia occurring in the FFBs are of a different composition than those in the ARBS. Also the feldspathic "prebasin" components of the FFBs have a lower average Mg/Fe ratio than the corresponding components of the ARBS; the average composition of the plagiociase in the FFBs is more sodic than that of the ARBS; and there are differences in relative abundances of rare earth elements. The two breccia types also have different provenances: the FFBs occur primarily in ejecta from North Ray crater and presumably derive from the Descartes Formation, while the ARBs are restricted to the Cayley plains. Together these observations suggest that although some type of fragmental breccia may have been a precursor to the ARBS, the FFBs of North Ray crater are not a significant component of the ARBs and, by inference, the Cayley plains. The average compositions of the prebasin components of the two types of fragmental breccia are generally similar to the composition of the feldspathic lunar meteorites. With 30-31% Al203, however, they are slightly richer in plagiociase than the most feldspathic lunar meteorites (approximately 29% Al203), implying that the crust of the early central nearside of the Moon contained a higher abundance of highly feldspathic anorthosite than typical lunar highlands, as inferred from the lunar meteorites. The ancient regolith breccias, as well as the current surface regolith ofthe Cayley plains, are more mafic than (1) prebasin regoliths in the Central Highlands and (2) regions of highlands presently distant from nearside basins because they contain a high abundance (approximately 30%) of mafic impact-melt breccias produced during the time of basin formation that is absent from other regoliths.

Korotev, Randy L.↗

DOE OSTI2026

Multiomics and deep learning dissect regulatory syntax in human development

Transcription factors establish cell identity during development by binding regulatory DNA in a sequence-specific manner, often promoting local chromatin accessibility and regulating gene expression1. Mapping accessible chromatin offers critical insights into transcriptional control, but available datasets for human development are restricted to bulk tissue, single organs or single modalities2. Here we present the Human Development Multiomic Atlas, a single-cell atlas of chromatin accessibility and gene expression from 817,740 fetal cells across 12 organs, spanning 203 cell types and more than 1 million candidate cis-regulatory elements, many of which exhibit organ-specific in vivo enhancer activity. Deep learning models trained to predict accessibility from local DNA sequence unravel a comprehensive lexicon of motifs that influence accessibility, including composite motifs exhibiting distinct syntactic constraints that are predicted to mediate transcription factor cooperativity. We identify ‘hard’ syntactic rules requiring precise motif spacing and orientation, ‘soft’ rules allowing flexible motif arrangements, and ubiquitous motifs inhibiting accessibility. Model-based interpretation of genetic variants reveals that disruption of motifs with positive and negative effects is associated with concordant effects on gene expression. Our work delineates how motif syntax governs cell-type-specific chromatin accessibility and provides a foundational resource for decoding cis-regulatory logic and interpreting genetic variation during human development.

59 BASIC BIOLOGICAL SCIENCES↗

NASA NTRS2002

Evolutionary Optimization of Quadrifilar Helical and Yagi-Uda Antennas

We present optimization results obtained for two type of antennas using evolutionary algorithms. A quadrifilar helical UHF antenna is currently flying aboard NASA's Mars Odyssey spacecraft and is due to reach final Martian orbit insertion in January, 2002. Using this antenna as a benchmark, we ran experiments employing a coevolutionary genetic algorithm to evolve a quadrifilar helical design in-situ - i.e., in the presence of a surrounding structure. Results show a 93% improvement at 400 MHz and a 48% improvement at 438 MHz in the average gain. The evolved antenna is also one-fourth the size. Yagi-Uda antennas are known to be difficult to design and optimize due to their sensitivity at high gain and the inclusion of numerous parasitic elements. Our fitness calculation allows the implicit relationship between power gain and sidelobe/backlobe loss to emerge naturally, a technique that is less complex than previous approaches. Our results include Yagi-Uda antennas that have excellent bandwidth and gain properties with very good impedance characteristics. Results exceeded previous Yagi-Uda antennas produced via evolutionary algorithms by at least 7.8% in mainlobe gain.

Lohn, Jason D.↗

DOE OSTI2023

A large sequenced mutant library – valuable reverse genetic resource that covers 98% of sorghum genes

SUMMARY Mutant populations are crucial for functional genomics and discovering novel traits for crop breeding. Sorghum , a drought and heat‐tolerant C4 species, requires a vast, large‐scale, annotated, and sequenced mutant resource to enhance crop improvement through functional genomics research. Here, we report a sorghum large‐scale sequenced mutant population with 9.5 million ethyl methane sulfonate (EMS)‐induced mutations that covered 98% of sorghum's annotated genes using inbred line BTx623. Remarkably, a total of 610 320 mutations within the promoter and enhancer regions of 18 000 and 11 790 genes, respectively, can be leveraged for novel research of cis ‐regulatory elements. A comparison of the distribution of mutations in the large‐scale mutant library and sorghum association panel (SAP) provides insights into the influence of selection. EMS‐induced mutations appeared to be random across different regions of the genome without significant enrichment in different sections of a gene, including the 5′ UTR, gene body, and 3′‐UTR. In contrast, there were low variation density in the coding and UTR regions in the SAP. Based on the K a / K s value, the mutant library (~1) experienced little selection, unlike the SAP (0.40), which has been strongly selected through breeding. All mutation data are publicly searchable through SorbMutDB ( https://www.depts.ttu.edu/igcast/sorbmutdb.php ) and SorghumBase ( https://sorghumbase.org/ ). This current large‐scale sequence‐indexed sorghum mutant population is a crucial resource that enriched the sorghum gene pool with novel diversity and a highly valuable tool for the Poaceae family, that will advance plant biology research and crop breeding.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2020

Bacterial genome editing by coupling Cre-lox and CRISPR-Cas9 systems

The past decade has been a golden age for microbiology, marked by the discovery of an unprecedented increase in the number of novel bacterial species. Yet gaining biological knowledge of those organisms has not kept pace with sequencing efforts. To unlock this genetic potential there is an urgent need for generic (i.e. non-species specific) genetic toolboxes. Recently, we developed a method, termed chassis-independent recombinase-assisted genome engineering (CRAGE), enabling the integration and expression of large complex gene clusters directly into the chromosomes of diverse bacteria. Here we expand upon this technology by incorporating CRISPR-Cas9 allowing precise genome editing across multiple bacterial species. To do that we have developed a landing pad that carries one wild-type and two mutant lox sites to allow integration of foreign DNA at two locations through Cre-lox recombinase-mediated cassette exchange (RMCE). The first RMCE event is to integrate the Cas9 and the DNA repair protein genes RecET, and the second RMCE event enables the integration of customized sgRNA and a repair template. Following this workflow, we achieved precise genome editing in four different gammaproteobacterial species. We also show that the inserted landing pad and the entire editing machinery can be removed scarlessly after editing. We report here the construction of a single landing pad transposon and demonstrate its functionality across multiple species. The modular design of the landing pad and accessory vectors allows design and assembly of genome editing platforms for other organisms in a similar way. We believe this approach will greatly expand the list of bacteria amenable to genetic manipulation and provides the means to advance our understanding of the microbial world.

59 BASIC BIOLOGICAL SCIENCES↗