CRISPR and Genetic Engineering

The paper develops fluorescently labeled split-QF and QF2 hemidrivers in zebrafish to simplify intersectional targeting and enable quality control of split-driver function.

This study defines the assembly pathway of a Tn7-like CAST, showing how DNA remodeling and TnsC positioning convert target recognition into directional transposition.

The paper identifies a phage-encoded small RNA that mimics the CRISPR-Cas9 guide RNA to inhibit the system and demonstrates its reprogrammability for DNA cleavage and trans-cleavage.

The paper shows Pseudomonas aeruginosa uses a CRISPR-Cas system to silence a proline-codon toxin that stops phage growth, revealing a defense mechanism against anti-CRISPR phages.

This paper introduces Shred-seq, a method to scan large-scale deletions in the human genome, revealing essential regions including noncoding sequences and establishing a lower bound for viability.

This paper identifies viral single-guide RNAs (vsgRNAs) that subvert bacterial CRISPR-Cas9 immunity by sequestering Cas9 and redirecting its activity, and shows they can enable genome editing in mammalian cells.

This paper demonstrates a gene drive strategy targeting KLHL10 in mice to increase homing efficiency by removing NHEJ-mutated sperm, but observes unexpected mutation absence in female offspring.

The paper introduces CIRI, a CRISPR-based platform for programmable cell fate conversion in hiPSCs, using single-cell genomics to identify regulatory factors for myogenic differentiation.

This paper introduces CRISPR-StAR, an in vivo CRISPR screening platform with improved normalization and analysis tools, showing it can identify tumor suppressor gene effects not observable in vitro.

This paper introduces a method using single-stranded DNA templates and bacteriophage RNA polymerases to produce functional RNAs in mammalian cells, demonstrating applications in CRISPR-based genome editing.

DropSynth-Gold uses Golden Gate assembly in emulsions to enable longer and more complex gene library synthesis with comparable fidelity to previous methods.

This study identifies two distinct RNA-guided transposition pathways used by IS110 transposons, resolving prior inconsistencies in their mechanism.

This study shows that base editing in human embryos produces efficient on-target changes without chromosomal abnormalities or large deletions, unlike CRISPR/Cas9-induced DSBs.

This paper develops inducible Calling Cards in mice to control transposon insertion timing in vivo, showing varying efficacy and developmental impacts based on the transcription factor used.

This study identifies a bacterial antiphage defense system combining Ago and SLFN domains, showing how ATP-driven tetramerization and RNA-guided filament assembly regulate nuclease activity.

This study compares non-viral CRISPR delivery methods in stem cells, finding that lipid-based delivery offers high efficiency but risks genomic instability, while CPPs reduce side effects at lower efficiency.

The paper improves a CRISPR-base editor tool for targeting virulence factors in Mycoplasma bovis by enabling multi-loci targeting and expanding targetable loci.

This study shows that CRISPR-based gene drives producing dominant female-sterile alleles can enhance population suppression by increasing elimination rates and reducing chasing dynamics.

This paper introduces NovoCAST, a new RNA-guided transposon system designed through computational protein design, enabling efficient and programmable DNA integration in human cells.

The paper develops a CRISPR/Cas9 method for editing Zymoseptoria tritici and shows that Mycgr3107904 contributes to disease progression.

This study shows that Cas12a2 preferentially cleaves collateral DNA over RNA, revealing the mechanism and implications for CRISPR-based immunity and technology.

FLInt 2.0 improves transgene integration in C. elegans by reducing false positives through targeted Cas9 cutting and fluorescence preservation during repair events.

The paper develops fully modified SpyCas9 guide RNAs that maintain or enhance genome editing efficacy in vitro and in vivo.

This paper introduces nickase-based MAGIC as a safer alternative to DSB-based CRISPR/Cas9 for generating genetic mosaics in Drosophila, showing that single-strand nicks can induce recombination with tunable efficiency.

This paper develops CRISPR-MACE, a platform for continuous directed evolution of CRISPR-Cas9 in human cells, generating variants with enhanced DNA binding and resistance to AcrIIA4.

This study shows that using multiple guide RNAs in CRISPR screens can reduce cell numbers needed while maintaining screening effectiveness.

This study reviews plant CRISPR studies and shows multiplex editing efficiency increases with more targets, suggesting feasibility of editing up to 100 genes in plants.

The paper introduces intcgRNA, a conditional guide RNA that uses endogenous intronic RNA to control CRISPR/Cas9 editing, showing improved specificity in human cells.

The paper introduces OMEGA, a low-cost method for assembling large gene libraries from oligopools, achieving high sequence recovery and uniformity.

This study solves cryo-EM structures of SpyCas9 bound to minicircle DNA, revealing how DNA shape affects Cas9 target interrogation and cleavage.