2026· Methods in molecular biology· Vol 3041, pp.
33-45
· 0 citations
Medicine
TL;DR
An improved tool called pSPIN-GG and supporting protocols for simplified CAST-based genome engineering are presented and refinements support accelerated library construction, reduce assembly and screening burden, and expand the accessibility of CAST systems for multiplexed bacterial genome engineering.
Overall, transposon-based tools represent a valuable and underexplored avenue to accelerate genome engineering and strain development in yeasts and filamentous fungi.
Bing-Yin Peng, Masahiro Tominaga, Cheng-Qiang Wang et al.· Yeast· 1 citation
An RNA-guided bridge recombinase system is engineered through rational mutagenesis and AI-assisted directed evolution, enabling programmable chromosomal rearrangements in both plant and mammalian cells and achieving up to a 29.8-fold increase in activity.
Rui Gao, Jingjing Wei, Chao Sun et al.· Trends in Biotechnology· 0 citations
Genomic manipulation has advanced from stochastic nuclease‐mediated disruption toward programmable, deterministic precision. Early clustered regularly interspaced short palindromic repeats (CRISPR) strategies enabled targeted mutagenesis through double‐strand breaks; however, their therapeutic application is limited by...
MORTISE is introduced, a compact Cas6f-based RNA system to repress target bacterial genes without relying on host RNA factors, enables multiplex control across bacteria, and supports pathway balancing when genetic knockouts are unsuitable.
This work compares the performance of multiple KRAB domain systems, develops an updated CRISPRi-specific on-target scoring scheme, and quantitatively characterize off-target effects associated with seed-sequence patterns.
Smriti Srikanth, Fengyi Zheng, Laura M Drepanos et al.· Cell Genomics· 0 citations
Summary Here, we present a CRISPR-Cas9 genome-editing protocol for Saccharomyces cerevisiae that simplifies guide construction and enables rapid guide iteration by replacing restriction/ligation cloning with PCR-based guide installation and seamless Cas9-plasmid recircularization. We describe single-guide RNA (sgRNA) a...
H. Rostamian, Ethan Madden, Frank M. Kaplan et al.· STAR Protocols· 0 citations
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