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    1. Developmental Biology
    2. Genetics and Genomics

    CRISPR/Cas9 and active genetics-based trans-species replacement of the endogenous Drosophila kni-L2 CRM reveals unexpected complexity

    Xiang-Ru Shannon Xu et al.
    Dissection of a cis-regulatory element (CRM) in its native chromosomal context using CRISPR/Cas9 editing and novel 'Active Genetics' reveals new features of CRM function and insights into how such regulatory elements change during evolution.
    1. Microbiology and Infectious Disease

    Exploiting CRISPR-Cas to manipulate Enterococcus faecalis populations

    Karthik Hullahalli et al.
    Conflicts between CRISPR-Cas systems and antibiotic resistance plasmids can be exploited to selectively eliminate antibiotic resistance from Enterococcus faecalis populations.
    1. Biochemistry and Chemical Biology

    CRISPR-Cas12a exploits R-loop asymmetry to form double-strand breaks

    Joshua C Cofsky et al.
    The DNA flanks on either side of an R-loop differ in stability, and CRISPR-Cas12 enzymes form double-strand breaks by targeting the more unstable flank with their single DNase active site.
    1. Microbiology and Infectious Disease

    Co-evolution within structured bacterial communities results in multiple expansion of CRISPR loci and enhanced immunity

    Nora C Pyenson, Luciano A Marraffini
    Cells must acquire multiple viral spacer sequences to neutralize mutant escaper phages and form colonies during the CRISPR-Cas immune response.
    1. Structural Biology and Molecular Biophysics

    Casposase structure and the mechanistic link between DNA transposition and spacer acquisition by CRISPR-Cas

    Alison B Hickman et al.
    The tetrameric structure of a casposase bound to DNA and its biochemical properties show how a transposase could have evolved to perform CRISPR-Cas spacer acquisition.
    1. Genetics and Genomics
    2. Neuroscience

    Dissection of central clock function in Drosophila through cell-specific CRISPR-mediated clock gene disruption

    Rebecca Delventhal et al.
    Through novel, cell-specific CRISPR tools to disrupt molecular clock genes, it was revealed that circadian rhythms are coordinated through a network, rather than by the clock of 'master regulatory' neurons.
    1. Developmental Biology

    Is it possible to reconstruct an accurate cell lineage using CRISPR recorders?

    Irepan Salvador-Martínez et al.
    Computer simulations reveal the potential and limitations of recently proposed CRISPR-based cell lineage recorders, and suggest how the recorders' design can be optimised to yield more accurate cell lineage trees.
    1. Biochemistry and Chemical Biology
    2. Structural Biology and Molecular Biophysics

    The Cas4-Cas1-Cas2 complex mediates precise prespacer processing during CRISPR adaptation

    Hayun Lee et al.
    During CRISPR adaptation, Cas4 forms a ternary complex with the Cas1-Cas2 spacer integration complex, an interaction that coordinates substrate hand-off following precise, PAM-dependent prespacer processing prior to integration.
    1. Biochemistry and Chemical Biology

    Structural basis for AcrVA4 inhibition of specific CRISPR-Cas12a

    Gavin J Knott et al.
    An ancestral helical bundle governs whether the crRNA-processing nuclease of CRISPR-Cas12a is exploited by a phage-encoded allosteric anti-CRISPR.
    1. Chromosomes and Gene Expression

    Synthetically modified guide RNA and donor DNA are a versatile platform for CRISPR-Cas9 engineering

    Kunwoo Lee et al.
    The guide RNA and donor DNA of the CRISPR/Cas system tolerate large chemical modifications and can be engineered for enhanced delivery and gene editing.