Biology subjects

Pandey, N.

Publications and source records attributed to Pandey, N..

At least 19 recordsLinked to original sources

bioRxiv2026

Structural basis of substrate recognition and conformational gating in the bacteriophage M15 metalloendopeptidase LysPH

The rapid emergence of antimicrobial resistance has led to a surge in multidrug-resistant strains, jeopardising the efficacy of frontline and last-resort antibiotics and thereby aggravating the antimicrobial resistance crisis. Bacteriophage-derived endolysins represent a promising class of next-generation antimicrobials. Here, we report the isolation of a novel bacteriophage, PA_Ganga_001, targeting multidrug-resistant Pseudomonas aeruginosa. Genomic sequencing of this phage identified a previously uncharacterised endolysin LysPH, a zinc-dependent globular endolysin that exhibits antibacterial activity against multiple multidrug-resistant Gram-negative pathogens, including Pseudomonas aeruginosa, Acinetobacter baumannii, and Klebsiella pneumoniae. Site-directed mutagenesis of the endolysin LysPH suggests that His77, Asp84, His159, Arg41, and Asp156 are essential for catalysis and substrate accommodation. To understand the structural basis of substrate binding and catalysis, we determined the crystal structures of the apo-enzyme (1.8 [A]) and the catalytically attenuated D156A mutant in complex with a synthetic pentapeptide stem (PGX) of peptidoglycan (2.0 [A]). The pentapeptide-bound complex revealed a defined substrate-binding groove, and a pronounced displacement of the Thr42-Ser75 loop was observed. This led to an approximately six-fold volumetric expansion of the catalytic cleft, indicative of a substrate-induced transition to an open, catalytically competent conformation. Notably, a distinctive C-terminal helical element, which diverges sequentially from characterised M15 homologues, is predicted to interact with the NAG-NAM scaffold and may contribute to the positioning of the stem pentapeptide for Zn2+ dependent catalysis. These distinct structural features establish the molecular basis for L-Ala-D-Glu hydrolysis by M15 bacteriophage endolysins and provide a foundational framework for the rational design and engineering of the next generation of antimicrobial enzymes.

biochemistry↗

bioRxiv2026

EnzFinder: a sustainable alternative to chemical synthesis

Enzymes have emerged as an important alternative to traditional catalysts in chemical industries over the past few decades owing to their sustainable nature. The application of enzymes in chemical synthesis relies on their ability to catalyse promiscuous reactions. Promiscuous activity of enzymes is an abundant phenomenon in nature; approximately 37% of Escherichia coli K12 enzymes show promiscuous activity. This highlights the vast expanse of chemical reactions that can be made biochemically feasible through selection of the correct candidate enzymes. Here, we present EnzFinder, a promiscuous enzyme prediction tool that filters candidate enzymes based on similarity in chemical transformation patterns and subsequently ranks them using substrate-product similarity, enabling enzyme prioritization up to the fourth level of EC classification without requiring sequence information. On a blind benchmarking set of 2,309 biochemical reactions, the method achieves substantially higher prediction accuracy than existing rule-based and deep-learning approaches, with improvements exceeding 20% at the sub-subclass level and significantly higher coverage at the fourth level. Application to industrially relevant reactions demonstrates EnzFinders ability to identify alternative enzymes with higher substrate similarity and improved kinetic potential. Furthermore, integration of EnzFinder with in silico retrosynthesis tools enables effective prioritization of enzymatic steps within hybrid chemical-biological pathways. Together, these results establish EnzFinder as a practical and interpretable tool for accelerating enzyme discovery and promoting greener, enzyme-driven synthesis routes.

synthetic biology↗

bioRxiv2025

Scalable flow synthesis of ultrasmall inorganic nanoparticles for biomedical applications via a confined impinging jet mixer

Ultrasmall inorganic nanoparticles (sub-5 nm) have unique biomedical advantages due to rapid clearance, enhanced imaging contrast, and potent therapeutic properties. However, current synthesis methods are limited by low throughput, polydispersity, and reliance on harsh conditions such as organic solvents or high temperatures. We report a scalable, single-step aqueous synthesis using a confined impinging jet mixer (CIJM) that produces size-controlled, clinically relevant nanoparticles, including silver sulfide, silver telluride, cerium oxide, and iron oxide, under ambient conditions. The resulting nanoparticles are homogeneous, stable, and preserve their functional biological properties. We demonstrate consistent performance across scales, establishing the CIJM as a versatile and reproducible method for producing ultrasmall inorganic nanoparticles suitable for clinical translation and high-throughput biomedical applications.

bioengineering↗

bioRxiv2025

Synergy unleashed: Leishmania donovani GP63 paralogues cooperatively function to drive parasite infectivity at various pathogenic stages and regulate host survival mechanisms

Highly plastic genome of Leishmania is thought to underlie its remarkable ability to adapt to diverse host microenvironments, yet how this plasticity translates into dermal versus visceral persistence remains unclear. GP63, a zinc metalloprotease, is a well-established determinant of promastigote infectivity, particularly in cutaneous leishmaniasis. Despite this, its role in amastigote stage and in visceral disease caused by Leishmania donovani remains undefined. Here, using an integrated approach combining phylogenetics, CRISPR-mediated genome editing, and biochemical analyses, we demonstrate that GP63 paralogues in L. donovani have undergone independent evolutionary diversification to support visceral infection. While GP63-paralogues encoded on chromosome 10, previously implicated in cutaneous manifestation caused by L. major, were found severely truncated and dispensable in L. donovani, two distinct GP63-paralogues encoded on chromosome 28 and 31 were found essential to establish visceral infection. LdGP63_31 facilitates promastigote attachment to macrophages by promoting lipid raft engagement and complement inactivation, thereby enabling host entry and subsequent amastigote differentiation. LdGP63_28 is essential for intracellular amastigote survival by suppressing host inflammatory pyroptosis. Structural and enzymatic analyses revealed preferential host localization and substrate specificities, possibly resulting from distinct amino acid substitution within conserved motifs of these proteases, indicating independent evolutionary adaptations. Importantly, loss of the LdGP63_28 more severely impaired amastigote reinfection than loss of its counterpart LdGP63_31. Together, these findings reveal coordinated functional specialization of GP63-paralogues ensuring visceral adaptation.

microbiology↗

bioRxiv2025

SAGA (Simplified Association Genomewide Analyses): a user-friendly Pipeline to Democratize Genome-Wide Association Studies

Genome-wide association studies (GWAS) have enabled clinicians and researchers to identify genetic variants linked to complex traits and diseases(1-3). However, GWAS still face several challenges, particularly regarding accessibility and reproducibility (4-6). Conducting these analyses often requires substantial bioinformatics expertise for data preprocessing, software installation, and scripting(7-10). We then developed SAGA ("Simplified Association Genome-wide Analyses"), a BASH-based, open-source, fully automated pipeline that integrates three widely adopted tools--PLINK(11), GMMAT(12), and SAIGE(13)--for accessible, robust, and reproducible GWAS. After installation, users simply need to provide genotype and phenotype files in standard formats. The pipeline automates preprocessing, association testing, and visualization, outputting summary statistics, Manhattan plots, and quantile-quantile plots. SAGA enables robust GWAS for users without scripting experience, expanding access to complex genetic analyses.

bioinformatics↗

bioRxiv2025

Estimation of stature from hand length and hand breadth in undergraduate medical students: An anthropometric study

BackgroundStature, a determinant that aids in personal identification, can be estimated using measurements of various body parts. This study measured the length and breadth of hands and the height of the participants, aiming to develop linear regression equations for stature estimation. MethodsThis is a cross-sectional descriptive study in which 195 medical students aged 18 to 24 were enrolled, and measurement of stature was done by stadiometer and hand length and breadth by vernier calliper, after an informed written consent was obtained. ResultsMean stature was higher in males (170.24{+/-}5.79 cm) as compared to females (159.01{+/-}7.261 cm). Mean length and breadth of both right and left hands were also found to be higher in males when compared with females. With a standard error of estimates of 5.083 cm and an R2 of 0.648, the multiple linear regression analysiss model summary concluded that R is .805, indicating a strong positive association between stature estimation and hand dimensions. ConclusionsHand length and hand breadth together may be a powerful predictor of stature, as indicated by the greater R value when compared to regression coefficients derived for other body parts. A variable entity, height is influenced by genetics and nutritional status. Future studies can be done taking these factors into consideration and also including a larger population size.

systems biology↗

bioRxiv2025

SKiD: A Structure-Oriented Kinetics Database of Enzyme-Substrate Interactions

Enzymes are essential biological catalysts that drive nearly all biochemical reactions. Understanding their efficiency and specificity involves studying enzyme kinetics, particularly the parameters kcat and Km. However, there is limited data linking these kinetic parameters with the three-dimensional (3D) structures of enzyme-substrate complexes. Since enzyme function is determined by its structure, such mapping enhances insight into structural basis of enzymatic function and supports applications in enzyme design, synthetic biology and metabolic engineering. To address this critical gap, this work presents SKiD (Structure-oriented Kinetics Database), a comprehensive, structured database integrating kcat and Km values with the corresponding 3D structural data. This is accomplished by integrating data from existing bioinformatics resources using automated programs to process the data and enhancing it with computational predictions. The erroneous data encountered during data integration is manually resolved. Metadata such as literature and assay conditions (e.g., pH and temperature) are preserved. The 3D coordinates of the modelled enzyme-substrate complexes are provided along with their UniProtKB identifier. The database is freely accessible from: https://doi.org/10.5281/zenodo.15355031.

bioinformatics↗

bioRxiv2025

Data-driven feedback augments ultrasound nanotheranostics in brain tumors

The blood-brain barrier (BBB) renders the delivery of nanomedicine in the brain ineffective and the detection of circulating disease-related DNA from the brain unreliable. Here, we show that the acoustic emission content of focused ultrasound-controlled microbubble dynamics (MB-FUS) incorporates precursor signals that allow large-data models to predict sonication regimens for safe and effective BBB opening. Crucially, closed-loop MB-FUS controller augmented by machine learning (ML-CL) expands the treatment window (4-fold), as compared to conventional controllers, by persistently and proactively maximizing the BBB permeability while preventing tissue damage. By successfully scaling up from mice to rats and from healthy to diseased brains (glioma), ML-CL rendered the BBB permeable to large nanoparticles and markedly improved the release and detection of tumor DNA in plasma. Together, our findings reveal the potential of data-driven feedback to support the development of next-generation AI-powered ultrasound systems for safe, robust, and efficient nanotheranostic targeting of brain diseases.

bioengineering↗

bioRxiv2025

Fighting Through the Heat: How Sexual Selection Influences Demography Under Recurrent Heatwaves

Sexual selection is a potent evolutionary force with complex effects. Strong sexual selection can enhance adaptation and reduce mutational load, while simultaneously reducing survival, or causing sexual conflict that reduces fitness for one or both sexes. Many populations today face not only gradual environmental changes but also extreme, short-term stress events like droughts or heatwaves. The combined effects of sexual and environmental selection on population demography during and after such events remain poorly understood, even though such combined effects could be crucial for the persistence of small, endangered populations under climate change. In this study, we investigated how sexual selection affects survival during environmental stress by manipulating the expression of an aggressive fighter morph in small populations of the male-dimorphic soil mite Sancassania berlesei, exposing some of these populations to recurrent periods of extreme heat and monitoring survival over eight generations. We found that heat exposure reduced survival, more severely in females than in males, and survival was lower in populations with higher fighter prevalence, but there was no interaction between temperature and fighter morph prevalence. Furthermore, survival declined across generations, and the decline was steeper in populations with lower prevalence of fighters, leading to the loss of their initial survival advantage by the last generation. T265625hree populations exposed to heat became extinct during the experiment, all from the reduced fighter expression treatment. Our findings imply that despite its cost to individual survival, sexual selection does not modulate population sensitivity to heatwaves over several generations. Furthermore, we demonstrate that these mortality costs of sexual selection are gradually compensated over successive generations, which could be a result of a more effective purging of inbreeding depression. Thus, while the additive effect of sexual selection and heatwaves on survival may increase demographic risks for bottlenecked populations in the short term, sexual selection may increase resilience of populations to prolonged bottlenecks.

evolutionary biology↗

bioRxiv2025

Understanding the biochemical significance of the mitochondrial pyruvate carrier in the rodent malarial parasite, Plasmodium berghei

Mitochondrial pyruvate carrier (MPC) protein has been established as the key pyruvate transporter across the inner mitochondrial membrane. In Plasmodium, flux through the tricarboxylic acid (TCA) cycle is low in the asexual stages that is upregulated in the gametocytes with greater utilization of glutamine than glucose to drive the cycle. Acetyl-CoA in the mitochondrion has been established to be essential for the parasite. In this context, we established the biochemical function of P. berghei MPC as a pyruvate transporter through functional complementation in yeast. In P. berghei, both subunits, MPC1 and MPC2 were found to localize to the mitochondrion and surprisingly, were found to be non-essential for the blood stages and for ookinete formation. Through metabolite tracing studies using 13C6-glucose and 13C515N2 L-glutamine, we have derived the biochemical basis for this observed phenotype of {Delta}mpc1{Delta}mpc2 parasites. Our studies suggest the presence of an aminotransferase localized to the mitochondrion that converts alanine to pyruvate, leading to the continued production of acetyl-CoA in the organelle even in the absence of MPC. This highlights the existence of metabolic plasticity, a survival strategy, that enables the utilization of different metabolic precursors for the generation of acetyl-CoA in the mitochondrion.

biochemistry↗

bioRxiv2024

The effects of Paternal Radiofrequency Radiation Exposure on Sperm and Testicular Integrity in F1-Male Swiss Albino Mice

The escalating prevalence of infertility and reproductive malignancies in the youth demographic warrants thorough investigation. The escalating ubiquity of communication devices, such as mobile phones and Wi-Fi, exposes individuals to unprecedented levels of Radiofrequency Radiation (RFR), raising concerns about its potential association with reproductive issues and cancer. In this study, we endeavored to elucidate the impact of RFR exposure on the fertility of F0 male Swiss albino mice, as well as the consequential effects on sperm quality and testicular morphology in F1 male mice during adulthood. F0 male mice were subjected to RFR exposure for 3 hours per day, twice daily, over a span of 35 days, followed by mating with naive, unexposed females, culminating in a comprehensive endpoint evaluation. The repercussions of RFR exposure included a diminution in sperm count, heightened incidence of sperm defects, augmented DNA damage in both sperm and bone marrow cells, diminished sperm viability, and aberrations in testicular histology. Subsequent mating of exposed male mice with naive females revealed no discernible effects on pregnancy outcomes, as indicated by the fertility index, litter size, and copulation index.Nevertheless, F1 male offspring exhibited a noteworthy reduction in sperm count and depletion of the germinal epithelium within seminiferous tubules in the testes. These findings propose that RFR exposure to F0 males may precipitate diminished sperm count in subsequent F1 generations, implicating the potential role of epigenetic modulation through the male germline.

cell biology↗

bioRxiv2024

CD14 Blockade Does Not Improve Outcomes of Deep Vein Thrombosis Following Inferior Vena Cava Stenosis in Mice

BackgroundNeutrophil-mediated persistent inflammation and neutrophil extracellular trap formation (NETosis) promote deep vein thrombosis (DVT). CD14, a co-receptor for toll-like receptor 4 (TLR4), is actively synthesized by neutrophils, and the CD14/TLR4 signaling pathway has been implicated in proinflammatory cytokine overproduction and several aspects of thromboinflammation. The role of CD14 in the pathogenesis of DVT remains unclear. ObjectiveTo determine whether CD14 blockade improves DVT outcomes. MethodsBulk RNA sequencing and proteomic analyses were performed using isolated neutrophils following inferior vena cava (IVC) stenosis in mice. DVT outcomes (IVC thrombus weight and length, thrombosis incidence, neutrophil recruitment, and NETosis) were evaluated following IVC stenosis in mice treated with a specific anti-CD14 antibody, 4C1, or control antibody. ResultsMice with IVC stenosis exhibited increased plasma levels of granulocyte colony-stimulating factor (G-CSF) along with a higher neutrophil-to-lymphocyte ratio and increased plasma levels of cell-free DNA, elastase, and myeloperoxidase. Quantitative measurement of total neutrophil mRNA and protein expression revealed distinct profiles in mice with IVC stenosis compared to mice with sham surgery. Neutrophils of mice with IVC stenosis exhibited increased inflammatory transcriptional and proteomic responses, along with increased expression of CD14. Treatment with a specific anti-CD14 antibody, 4C1, did not result in any significant changes in the IVC thrombus weight, thrombosis incidence, or neutrophil recruitment to the thrombus. ConclusionThe results of the current study are important for understanding the role of CD14 in the regulation of DVT and suggest that CD14 lacks an essential role in the pathogenesis of DVT following IVC stenosis.

pathology↗

bioRxiv2023

An explainable model using Graph-Wavelet for predicting biophysical properties of proteins and measuring mutational effects

Proteins hold multispectral patterns of different kinds of physicochemical features of amino acids in their structures, which can help understand proteins behavior. Here, we propose a method based on the graph-wavelet transform of signals of features of amino acids in protein residue networks derived from their structures to achieve their abstract numerical representations. Such abstract representations of protein structures hand in hand with amino-acid features can be used for different purposes, such as modelling the biophysical property of proteins. Our method outperformed graph-Fourier and convolutional neural-network-based methods in predicting the biophysical properties of proteins. Even though our method does not predict deleterious mutations, it can summarize the effect of an amino acid based on its location and neighbourhood in protein-structure using graph-wavelet to estimate its influence on the biophysical property of proteins. Such an estimate of the influence of amino-acid has the potential to explain the mechanism of the effect of deleterious non-synonymous mutations. Thus, our approach can reveal patterns of distribution of amino-acid properties in the structure of the protein in the context of a biophysical property for better classification and more insightful understanding.

bioinformatics↗

bioRxiv2023

Assessment of Nutrient Composition and utilization of some popular wild Edible fruits of Kumaun Himalayan Region as Anti-oxidant agents

Food and Nutrition security is the main problem faced by developing and underdeveloped countries. Proper utilization of wild edible plants for nutrition security is a great choice for reducing the wastage of these powerful foods. The present investigation has been made to understand the nutritional and phytochemical parameter of selected underutilized fruits such as Diospyros kaki, Pyrus pashia, Ficus semicordata, Diploknema butyracea, Pyracantha crenulata, and Rubus niveus. The proximate analyses showed that Diospyros kaki is most promising fruit with 44.86{+/-} 0.4 mg/g carbohydrate, 9.29{+/-}0.80 mg/g protein, 81.31{+/-}0.4, 7% moisture and 1.1{+/-}0.11% little amount of ash content. Micronutrient iron was quantify highest in Pyracantha crenulata (3.52{+/-}0.24mg/100g) and zinc in Rubus niveus(8.13{+/-}0.05 mg/100g). Phytochemical screening was recorded in the ethanolic and aqueous extract, in which Phenolic and flavonoid content were highest in ethanolic extract of Rubus niveus; 64.05{+/-}0.13mg GAE/gm, 108.83{+/-}2.93mg QE/ g extract respectively and tannin content was highest in case of Diospyros kaki (79.94{+/-}0.40mg TAE/100g of extract). The free radical scavenging activity of fruits have been analyzed by DPPH, H2O2 and NO assay; it was observed that the ethanolic extract of Rubus niveus fruit is most promising with an IC50value 16.97g/ml. In contrast, aqueous extract scored IC50value 28.86g/ml. The lowest IC50 value was found in aqueous extract of Diospyros kaki i.e., 81.9 g/ml in DPPH assay. The potential usage of these plants will help the people of these regions to combat nutrient deficiency diseases. Further development of research may help us to come out with powerful functional foods using these fruits. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=136 SRC="FIGDIR/small/545978v1_ufig1.gif" ALT="Figure 1"> View larger version (34K): org.highwire.dtl.DTLVardef@11d5ddborg.highwire.dtl.DTLVardef@fce3cborg.highwire.dtl.DTLVardef@cf5f67org.highwire.dtl.DTLVardef@808054_HPS_FORMAT_FIGEXP M_FIG C_FIG

biochemistry↗

bioRxiv2023

Centromere Innovations Within a Mouse Species

Mammalian centromeres direct faithful genetic inheritance and are typically characterized by regions of highly repetitive and rapidly evolving DNA. We focused on a mouse species, Mus pahari, that we found has evolved to house centromere-specifying CENP-A nucleosomes at the nexus of a satellite repeat that we identified and term {pi}-satellite ({pi}-sat), a small number of recruitment sites for CENP-B, and short stretches of perfect telomere repeats. One M. pahari chromosome, however, houses a radically divergent centromere harboring [~]6 Mbp of a homogenized {pi}-sat-related repeat, {pi}-satB, that contains >20,000 functional CENP-B boxes. There, CENP-B abundance drives accumulation of microtubule-binding components of the kinetochore, as well as a microtubule-destabilizing kinesin of the inner centromere. The balance of pro and anti-microtubule-binding by the new centromere permits it to segregate during cell division with high fidelity alongside the older ones whose sequence creates a markedly different molecular composition. TeaserChromatin and kinetochore alterations arise in response to evolutionarily rapid changes to underlying repetitive centromere DNA.

genomics↗

bioRxiv2023

The HRA Organ Gallery Affords Immersive Superpowers for Building and Exploring the Human Reference Atlas with Virtual Reality

The Human Reference Atlas (HRA, https://humanatlas.io) funded by the NIH Human Biomolecular Atlas Program (HuBMAP, https://commonfund.nih.gov/hubmap) and other projects engages 17 international consortia to create a spatial reference of the healthy adult human body at single-cell resolution. The specimen, biological structure, and spatial data that define the HRA are disparate in nature and benefit from a visually explicit method of data integration. Virtual reality (VR) offers unique means to enable users to explore complex data structures in a threedimensional (3D) immersive environment. On a 2D desktop application, the 3D spatiality and real-world size of the 3D reference organs of the atlas is hard to understand. If viewed in VR, the spatiality of the organs and tissue blocks mapped to the HRA can be explored in their true size and in a way that goes beyond traditional 2D user interfaces. Added 2D and 3D visualizations can then provide data-rich context. In this paper, we present the HRA Organ Gallery, a VR application to explore the atlas in an integrated VR environment. Presently, the HRA Organ Gallery features 55 3D reference organs,1,203 mapped tissue blocks from 292 demographically diverse donors and 15 providers that link to 5,000+ datasets; it also features prototype visualizations of cell type distributions and 3D protein structures. We outline our plans to support two biological use cases: on-ramping novice and expert users to HuBMAP data available via the Data Portal (https://portal.hubmapconsortium.org), and quality assurance/quality control (QA/QC) for HRA data providers. Code and onboarding materials are available at https://github.com/cns-iu/ccf-organ-vr-gallery#readme.

bioinformatics↗

bioRxiv2023

Deciphering the phenotypic heterogeneity and drug response in cancer cells using genome-wide activity and interaction of chromatin domains.

The effect of co-localization of genes in the topologically associated domains (TADs) and their activity as a regulatory unit in cancer samples and cells, together with drug-response, needs comprehensive analysis. Here, we analyzed the activity of TADs using cancer-cell transcriptomes along with chromatin-interaction and epigenome profiles to understand their relationship with drug-response. Our analysis of 819 cancer cell-line transcriptomes revealed that their response to multiple drugs was more correlated with the activity of individual TADs than genes. Applying our approach to 9014 cancer patients data (20 different cancer types) also revealed a higher association between survival and the activity of thousands of individual TADs in comparison to their genes. CRISPR-mediated knock-out of regulatory sites inside a TAD associated with cisplatin-response of oral cancer cells and discovery of primate-specific gain of synteny of genes within a TAD containing EGFR gene and its contribution towards cancer malignancy demonstrate greater utility of TAD-activity based analysis.

cancer biology↗

bioRxiv2022

CRISPRi in Deinococcus radiodurans

The extremely radiation resistant bacterium, Deinococcus radiodurans, is a microbe of importance, both, for studying stress tolerance mechanisms and as a chassis for industrial biotechnology. However, the molecular tools available for use in this organism continue to be limiting. In view of this, the CRISPR-Cas tools provide a large repertoire of applications for gene manipulation. We show the utility of the type I-E Cascade system for knocking down gene expression in this organism. A single-vector system was designed for expression of the Cascade components as well as the crRNA. The type I-E Cascade system was better tolerated than the type II-A Cas9 system in D. radiodurans. An assayable acid phosphatase gene, phoN integrated into the genome of this organism could be knocked down to 10% of its activity using the Cascade system. Cascade-based knockdown of ssb, a gene important for radiation resistance resulted in poor recovery post irradiation. Targeting the Radiation and Desiccation Resistance Motif (RDRM), upstream of the ssb, prevented de-repression of its expression upon radiation exposure. In addition to this, multi-locus targeting was demonstrated on the deinococcal genome, by knocking down both phoN and ssb expression simultaneously. The programmable CRISPRi tool developed in this study will facilitate study of essential genes, hypothetical genes, cis-elements involved in radiation response as well as enable metabolic engineering in this organism. Further the tool is amenable for implementing high-throughput approaches for such studies.

molecular biology↗