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Wang et al. developed a transformer base editor system in which the enzyme activity of the base editor is turned on only at the on-target site, therefore minimizing genome-wide and transcriptome-wide off-target mutations.
Zhang et al. design optogenetically controlled artificial transport vehicles that can be activated reversibly to manipulate cargo transport, impede neurite development and functionally characterize filopodial networks in axolotls.
Gao et al. developed a CRISPR–Cas9-based system in which sgRNA production is controlled by the endogenous promoter to monitor the expression of weakly expressed genes and long non-coding RNAs in mammalian cells.
Moghadam et al. developed a CRISPR transcriptional repressor to silence MyD88 expression in vivo to modulate immune response against AAV gene therapy and septicaemia.
Massou et al. combine cell stretching, super-resolution imaging and single-protein tracking to investigate integrin-based mechanosensing in live cells.
Li and colleagues report base editor variants with improved targeting efficiencies and broader editing windows by fusing the original base editors with the single-stranded DNA-binding domain of Rad51.
Artegiani, Hendriks et al. describe a CRISPR–Cas9-based method to efficiently generate human knock-in organoids using non-homologous end joining to study rare intestinal cell types and human hepatocyte division.
Yue, Zong, Li, Li, Zhang, Wu et al. introduce an in toto live-imaging system to track cardiac ventricle chamber formation at single-cell resolution for up to 1.5 days and digitally reconstruct cell dynamics.
Kim et al. develop an optogenetic visualization approach that can rapidly and reversibly trap messenger RNA molecules in protein clusters, thereby restricting their access to ribosomes and dampening translation efficiency.
He and colleagues develop itChIP-seq based on simultaneous cellular indexing and chromatin tagmentation. itChIP-seq is applicable to both low-input and single-cell analyses of chromatin states.
Simunovic et al. use human embryonic stem cells to generate a three-dimensional model of a human pre-gastrulation epiblast and show that anterior–posterior symmetry breaking can be induced by BMP4 and WNT signalling.
Coppé and colleagues design a peptide phosphorylation-screening system that simultaneously measures the enzymatic activity of multiple kinases, identifying mechanisms of therapy resistance and druggable targets in colorectal cancer and melanoma.
Gutnick et al. design a light-sensitive small molecule, zapalog, which reversibly dimerizes any two proteins. Tethering mitochondria to active kinesin motors uncovers distinct modes of mitochondrial motility in axons.
Wang et al. developed an inducible CRISPR–Cas9 system, in which guide RNA release is controlled by specific microRNAs, and demonstrated its application as a microRNA sensor and cell-type-specific genome regulator.
Nair et al. report the generation of human ESC-derived mature and functional β cells in vitro with a culture system including a step to induce clustering of immature β-like cells.
Giulitti et al. deliver modified mRNAs encoding OCT3/4, SOX2, KLF4 and cMYC as well as NANOG in microfluidics to directly convert human fibroblasts into naive induced pluripotent stem cells; the confined environment leads to enhanced efficiency and homogeneity compared to traditional methods.
Harada et al. develop a chromatin integration labelling (ChIL) method to map distributions of histone modifications and DNA-binding factors at low-input or even single-cell levels.
Li and colleagues develop a CRISPR–Cas9-based screen strategy that combines base editing and haploid embryonic stem cell technologies to identify amino acids critical for protein function in mice.
Mehta et al. create single-fluorophore kinase activity sensors for PKA, PKC and ERK in different colours, and demonstrate that they enable multiplex imaging of a combination of signalling activities in cell lines and primary rat neurons.
Sozen et al. devise an approach to combine embryonic stem cells, trophoblast stem cells and extra-embryonic endoderm stem cells into self-assembling embryo-like structures, which recapitulate key hallmarks of gastrulation in vitro.