Biological techniques

  • Article
    | Open Access

    Schinzel-Giedion syndrome (SGS) is a fatal developmental syndrome characterized by severe intellectual and physical deficits due, at least in part, to early neurodegeneration. Here the authors introduce a human SGS model that displays disease-relevant phenotypes to demonstrate that neuronal death in SGS originates from developmental alterations mainly in safeguarding cell identity and homeostasis.

    • Federica Banfi
    • , Alicia Rubio
    •  & Alessandro Sessa
  • Article
    | Open Access

    STAT1a is required for pro-inflammatory responses in macrophages. Here the authors reveal that post-translational modification of STAT1a, ADPribosylation, plays a critical role in enhancer formation and activation, thus modulating IFNγ-stimulated inflammatory responses in macrophages.

    • Rebecca Gupte
    • , Tulip Nandu
    •  & W. Lee Kraus
  • Article
    | Open Access

    Pyrrolysine (Pyl) exists in nature as the 22nd proteinogenic amino acid, but studies of Pyl have been hindered by the difficulty and inefficiency of both its chemical and biological syntheses. Here, the authors developed an improved PANCE approach to evolve the pylBCD pathway for increased production of Pyl proteins in E. coli.

    • Joanne M. L. Ho
    • , Corwin A. Miller
    •  & Matthew R. Bennett
  • Article
    | Open Access

    Secondary structures and long-range RNA interactions of the SARS-CoV-2 genome have been investigated by various sequencing methods. Here the authors use an RNA-RNA hybrid sequencing method to predict the secondary and tertiary structure of the SRAS-CoV-2 RNA genome in the virion.

    • Changchang Cao
    • , Zhaokui Cai
    •  & Yuanchao Xue
  • Article
    | Open Access

    The current biorefineries yield lignin with inadequate fractionation for bioconversion, yet substantial changes of these biorefinery designs could jeopardize carbohydrate efficiency and increase capital costs. Here the authors resolve the dilemma by designing ‘plug-in processes of lignin’ to enable economic waste valorization.

    • Zhi-Hua Liu
    • , Naijia Hao
    •  & Joshua S. Yuan
  • Article
    | Open Access

    Human Treg cells are central to immune tolerance, yet their heterogeneity and differentiation remain incompletely understood. Here the authors perform single-cell RNA and T cell receptor sequencing to resolve Treg cells from healthy individuals and patients with or without acute graft-versus-host disease revealing Treg complexity in health and disease.

    • Yuechen Luo
    • , Changlu Xu
    •  & Xiaoming Feng
  • Article
    | Open Access

    Current near-IR optogenetic systems to regulate transcription consist of a number of large protein components. Here the authors report a smaller single-component near-IR system, iLight, developed from a bacterial phytochrome that they use to control gene transcription in bacterial and mammalian cells.

    • Andrii A. Kaberniuk
    • , Mikhail Baloban
    •  & Vladislav V. Verkhusha
  • Article
    | Open Access

    The electron-withdrawing target (EWT)-induced fluorescence quenching is an unsolved issue in intramolecular charge transfer (ICT) fluorophores that limits their applicability. Here, the authors report a simple and generalizable strategy to reverse the EWT-induced quenching mode into light-up mode, by introducing an indazole building block between the π-bridge and the donor in the ICT scaffold.

    • Chenxu Yan
    • , Zhiqian Guo
    •  & Wei-Hong Zhu
  • Review Article
    | Open Access

    This review presents an overview of scenarios where van der Waals (vdW) materials provide unique advantages for nanophotonic biosensing applications. The authors discuss basic sensing principles based on vdW materials, advantages of the reduced dimensionality as well as technological challenges.

    • Sang-Hyun Oh
    • , Hatice Altug
    •  & Michael S. Strano
  • Article
    | Open Access

    Precise and dynamic manipulation of nano-objects on a large scale has been challenging. Here, the authors introduce acoustoelectronic nanotweezers, combining precision of electronic tweezers with large-field dynamic control of acoustic tweezers, demonstrating complex patterning of sub-100 nm objects.

    • Peiran Zhang
    • , Joseph Rufo
    •  & Tony Jun Huang
  • Article
    | Open Access

    Applying complexome profiling, Evers et al. unravel the composition of mitochondrial oxidative phosphorylation complexes in P. falciparum asexual and sexual blood stages. Abundance of these complexes differs between both stages, supporting the hypothesis that a mitochondrial metabolic switch is central to gametocyte development and functioning.

    • Felix Evers
    • , Alfredo Cabrera-Orefice
    •  & Taco W. A. Kooij
  • Article
    | Open Access

    Many proteins exist in various proteoforms but detecting these variants by bottom-up proteomics remains difficult. Here, the authors present a computational approach based on peptide correlation analysis to identify and characterize proteoforms from bottom-up proteomics data.

    • Isabell Bludau
    • , Max Frank
    •  & Ruedi Aebersold
  • Article
    | Open Access

    Particle fusion can improve signal-to-noise ratio in single molecule localization microscopy, but is limited by structural heterogeneity. Here, the authors demonstrate an unsupervised classification method that differentiates structurally different DNA origami structures without prior knowledge.

    • Teun A.P.M. Huijben
    • , Hamidreza Heydarian
    •  & Bernd Rieger
  • Article
    | Open Access

    Fat synthesis is necessary for normal physiology, but its dysregulation contributes to the pathology of many diseases. Here, the authors report a high-resolution mass spectrometry approach that quantifies fat synthesis flux in humans and mice following a brief and low dose of deuterated water.

    • Xiaorong Fu
    • , Stanisław Deja
    •  & Shawn C. Burgess
  • Article
    | Open Access

    Super-resolution microscopy is often limited by photobleaching or uneven distribution of fluorophores. The authors present a label-free superresolution method termed VISTA, combining sample-expansion and vibrational imaging, with resolution down to 78 nm in protein-rich biological structures in cells and tissues.

    • Chenxi Qian
    • , Kun Miao
    •  & Lu Wei
  • Article
    | Open Access

    Here, Geoghegan, Evelyn et al. provide a lattice light-sheet microscopy based 4D imaging pipeline to quantitatively investigate Plasmodium spp. invasion and show that the nascent parasitophorous vacuole is predominantly formed from host’s erythrocyte membrane and undergoes continuous remodeling throughout invasion.

    • Niall D. Geoghegan
    • , Cindy Evelyn
    •  & Kelly L. Rogers
  • Article
    | Open Access

    Voltage-sensitive dye imaging (VSDI) is a powerful technique for measuring membrane potential dynamics of neurons but the effective resolution is limited. Here, the authors developed an in silico model of VSDI to probe activity in a biologically detailed reconstruction of rodent neocortical microcircuits.

    • Taylor H. Newton
    • , Michael W. Reimann
    •  & Henry Markram
  • Article
    | Open Access

    Directed evolution commonly relies on point mutations but InDels frequently occur in evolution. Here the authors report a protein-engineering framework based on InDel mutagenesis and fragment transplantation resulting in greater catalysis and longer glow-type bioluminescence of the ancestral luciferase.

    • Andrea Schenkmayerova
    • , Gaspar P. Pinto
    •  & Jiri Damborsky
  • Article
    | Open Access

    Bone marrow chimaeric mice are a valuable tool in research, but require myeloablative conditioning. Here the authors demonstrate efficient FACS-free enrichment of haematopoietic stem and progenitor cells for transplantation into unconditioned recipient mice, as well as for genetic engineering using polyvinyl alcohol based media.

    • Kiyosumi Ochi
    • , Maiko Morita
    •  & Satoshi Yamazaki
  • Article
    | Open Access

    Mobile element insertions (MEIs) are a source of repetitive genetic variation and can lead to genetic disorders. Here the authors use Cas9-targeted nanopore sequencing to efficiently saturate enrichment for known and non-reference MEIs.

    • Torrin L. McDonald
    • , Weichen Zhou
    •  & Alan P. Boyle
  • Article
    | Open Access

    Formalin-fixed, paraffin-embedded (FFPE) specimens are an attractive resource for retrospective clinical proteomics studies. Here, the authors benchmark and optimize proteomics workflows for FFPE sample analysis and provide guidelines on which methods to use for different samples and applications.

    • Corinna Friedrich
    • , Simon Schallenberg
    •  & Philipp Mertins
  • Article
    | Open Access

    Inaccurate cell segmentation has been the major problem for cell-type identification and tissue characterization of the in situ spatially resolved transcriptomics data. Here we show a robust cell segmentation-free computational framework (SSAM), for identifying cell types and tissue domains in 2D and 3D.

    • Jeongbin Park
    • , Wonyl Choi
    •  & Naveed Ishaque
  • Article
    | Open Access

    Technical advancements have significantly improved early diagnosis of cervical cancer, but accurate diagnosis is still difficult due to various practical factors. Here, the authors develop an artificial intelligence assistive diagnostic solution to improve cervical liquid-based thin-layer cell smear diagnosis according to clinical TBS criteria in a large multicenter study.

    • Xiaohui Zhu
    • , Xiaoming Li
    •  & Yanqing Ding
  • Article
    | Open Access

    Cyanobacterial thylakoid membranes host the molecular machinery for the light-dependent reactions of photosynthesis and respiratory electron flow. Here, the authors show that newly synthesized thylakoids emerge between the plasma membrane and pre-existing thylakoids and describe the time-dependent assembly process of photosynthetic complexes.

    • Tuomas Huokko
    • , Tao Ni
    •  & Lu-Ning Liu
  • Article
    | Open Access

    The epigenetic mechanisms coordinating the maintenance of adult cellular lineages remain poorly understood. Here the authors demonstrate that HIRA, a H3.3 histone chaperone, establishes the chromatin landscape required for skeletal muscle cell identity.

    • Joana Esteves de Lima
    • , Reem Bou Akar
    •  & Frédéric Relaix
  • Article
    | Open Access

    The effect of fallopian tube’s curvature on sperm motion has not been studied in detail. Here, the authors use droplet microfluidics to create soft curved interfaces, revealing a dynamic switch in sperm motility from a progressive surface-aligned mode at low curvatures, to an aggressive surface-attacking mode at high curvatures.

    • Mohammad Reza Raveshi
    • , Melati S. Abdul Halim
    •  & Reza Nosrati
  • Article
    | Open Access

    Several existing algorithms predict the methylation of DNA using Nanopore sequencing signals, but it is unclear how they compare in performance. Here, the authors benchmark the performance of several such tools, and propose METEORE, a consensus tool that improves prediction accuracy.

    • Zaka Wing-Sze Yuen
    • , Akanksha Srivastava
    •  & Eduardo Eyras
  • Article
    | Open Access

    Early stages of embryogenesis are known to depend on subcellular localization and transport of maternal mRNA, but systematic analyses have been hindered by a lack of methods for tracking of RNA. Here the authors combine spatially-resolved transcriptomics and single-cell RNA labeling to perform a spatio-temporal analysis of the transcriptome during early zebrafish development, revealing insights into this process.

    • Karoline Holler
    • , Anika Neuschulz
    •  & Jan Philipp Junker
  • Article
    | Open Access

    Currently relatively few functional probes for Raman-based live-cell profiling exist. Here the authors build on their previous ultra-bright Raman dots to devise a 14-plexed Raman probe panel to quantify cell surface proteins, endocytosis activities and metabolic dynamics of single live cells.

    • Chen Chen
    • , Zhilun Zhao
    •  & Wei Min
  • Article
    | Open Access

    Here the authors use RT-qPCR and mass spectrometry to analyze longitudinal blood samples from intensive care unit (ICU) COVID-19 patients and controls. They find that viral RNA and pentraxin-3 predict 28-day ICU mortality and that galectin-3-binding protein is an interaction partner of SARS-CoV-2 spike glycoprotein with antiviral properties.

    • Clemens Gutmann
    • , Kaloyan Takov
    •  & Manuel Mayr
  • Article
    | Open Access

    The identification of HLA peptides by mass spectrometry is non-trivial. Here, the authors extended and used the wealth of data from the ProteomeTools project to improve the prediction of non-tryptic peptides using deep learning, and show their approach enables a variety of immunological discoveries.

    • Mathias Wilhelm
    • , Daniel P. Zolg
    •  & Bernhard Kuster