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Prime editors are reverse transcriptase (RT)-based genome-editing tools that utilize double-strand break (DSB)-free mechanisms to decrease off-target editing in genomes and enhance the efficiency of targeted insertions. The multiple prime editors that have been developed within a short span of time are a testament to the potential of this technique for targeted insertions. This is mainly because of the possibility of generation of all types of mutations including deletions, insertions, transitions, and transversions. Prime editing reverses several bottlenecks of gene editing technologies that limit the biotechnological applicability to produce designer crops. This review evaluates the status and evolution of the prime editing technique in terms of the types of editors available up to prime editor 5 and twin prime editors, and considers the developments in plants in a systematic manner. The various factors affecting prime editing efficiency in plants are discussed in detail, including the effects of temperature, the prime editing guide (peg)RNA, and RT template amongst others. We discuss the current obstructions, key challenges, and available resolutions associated with the technique, and consider future directions and further improvements that are feasible to elevate the efficiency in plants.

Plant biotechnology is rife with new advances in transformation and genome engineering techniques. A common requirement for delivery and coordinated expression in plant cells, however, places the design and assembly of transformation constructs at a crucial juncture as desired reagent suites grow more complex. Modular cloning principles have simplified some aspects of vector design, yet many important components remain unavailable or poorly adapted for rapid implementation in biotechnology research. Here, we describe a universal Golden Gate cloning toolkit for vector construction. The toolkit chassis is compatible with the widely accepted Phytobrick standard for genetic parts, and supports assembly of arbitrarily complex T‐DNAs through improved capacity, positional flexibility, and extensibility in comparison to extant kits. We also provision a substantial library of newly adapted Phytobricks, including regulatory elements for monocot and dicot gene expression, and coding sequences for genes of interest such as reporters, developmental regulators, and site‐specific recombinases. Finally, we use a series of dual‐luciferase assays to measure contributions to expression from promoters, terminators, and from cross‐cassette interactions attributable to enhancer elements in certain promoters. Taken together, these publicly available cloning resources can greatly accelerate the testing and deployment of new tools for plant engineering.

The advancement in high power lasers has urged the requisite of efficient optical limiting materials for both eye and sensor protection. The discovery of atomically thin 2D transition metal dichacogenides with distinctive properties has paved the way for a variety of applications including optical limiting. Until recently, the optical limiting effect exhibited by 2D materials is inferior to the benchmark materials fullerene (C₆₀) and graphene. This article reports the optical limiting activity of the 2D transition metal dichalcogenide, titanium disulfide (TiS₂) nanosheets, using optical and photoacoustic z‐scan techniques. The 77% nonlinear optical limiting exhibited by the TiS₂sheets with 73% linear‐transmittance is superior to that of any other existing 2D dichalcogenide sheets, graphene, and the benchmark optical limiting material, C₆₀. The enhanced nonlinear response is attributed to the concerted effect of 2‐photon and the induced excited state absorptions. By using photoacoustic z‐scan, a unique tool developed to determine the nonlinear optical limiting mechanism in materials, it is found that the optical limiting exhibited by TiS₂2D sheets and graphene are mainly due to nonlinear absorption rather than scattering effects. These results have opened the door for 2D‐dichalcogenide‐materials‐based highly efficient optical limiters, especially at low fluences.