Cas12a-mediated gene targeting by sequential transformation strategy in Arabidopsis thaliana.

Gene targeting (GT) allows precise manipulation of genome sequences, such as knock-ins and sequence substitutions, but GT in seed plants remains a challenging task. Engineered sequence-specific nucleases (SSNs) are known to facilitate GT via homology-directed repair (HDR) in organisms. Here, we demonstrate that Cas12a and a temperature-tolerant Cas12a variant (ttCas12a) can efficiently establish precise and heritable GT at two loci in Arabidopsis thaliana (Arabidopsis) through a sequential transformation strategy.

Application of multiple sgRNAs boosts efficiency of CRISPR/Cas9-mediated gene targeting in Arabidopsis.

Background: Precise gene targeting (GT) is a powerful tool for heritable precision genome engineering, enabling knock-in or replacement of the endogenous sequence via homologous recombination. We recently established a CRISPR/Cas9-mediated approach for heritable GT in Arabidopsis thaliana (Arabidopsis) and rice and reported that the double-strand breaks (DSBs) frequency of Cas9 influences the GT efficiency. However, the relationship between DSBs and GT at the same locus was not examined.

Chromosome-scale assembly and gene editing of Solanum americanum genome reveals the basis for thermotolerance and fruit anthocyanin composition.

Solanum americanum serves as a promising source of resistance genes against potato late blight and is considered as a leafy vegetable for complementary food and nutrition. The limited availability of high-quality genome assemblies and gene annotations has hindered the exploration and exploitation of stress-resistance genes in S. americanum.

Two florigens and a florigen-like protein form a triple regulatory module at the shoot apical meristem to promote reproductive transitions in rice.

Many plant species monitor and respond to changes in day length (photoperiod) for aligning reproduction with a favourable season. Day length is measured in leaves and, when appropriate, leads to the production of floral stimuli called florigens that are transmitted to the shoot apical meristem to initiate inflorescence development. Rice possesses two florigens encoded by HEADING DATE 3a (Hd3a) and RICE FLOWERING LOCUS T 1 (RFT1).

Precise and heritable gene targeting in rice using a sequential transformation strategy.

Gene targeting (GT) is a powerful tool for modifying endogenous genomic sequences of interest, such as sequence replacement and gene knockin. Although the efficiency of GT is extremely low in higher plants, engineered sequence-specific nucleases (SSNs)-mediated double-strand breaks (DSBs) can improve GT frequency. We recently reported a CRISPR-Cas9-mediated approach for heritable GT in , called the "sequential transformation" strategy.

The Sm core protein SmEb regulates salt stress responses through maintaining proper splicing of RCD1 pre-mRNA in Arabidopsis.

Salt stress adversely impacts crop production. Several spliceosome components have been implicated in regulating salt stress responses in plants, however, the underlying molecular basis is still unclear. Here we report that the spliceosomal core protein SmEb is essential to salt tolerance in Arabidopsis.

Natural variation in promoter hinders salt resistance during tomato domestication.

Increasing soil salinization seriously impairs plant growth and development, resulting in crop loss. The Salt-Overly-Sensitive (SOS) pathway is indispensable to the mitigation of Na toxicity in plants under high salinity. However, whether natural variations of contribute to salt tolerance has not been reported.

Insights into the molecular mechanisms of CRISPR/Cas9-mediated gene targeting at multiple loci in Arabidopsis.

Homologous recombination-mediated gene targeting (GT) enables precise sequence knockin or sequence replacement, and thus is a powerful tool for heritable precision genome engineering. We recently established a clustered regularly interspaced short palindromic repeats/clustered regularly interspaced short palindromic repeats-associated protein 9 (CRISPR/Cas9)-mediated approach for heritable GT in Arabidopsis (Arabidopsis thaliana), but its broad utility was not tested, and the underlying molecular mechanism was unclear. Here, we achieved precise GT at 14 out of 27 tested endogenous target loci using the sequential transformation approach and obtained vector-free GT plants by backcrossing.

Gene Targeting Facilitated by Engineered Sequence-Specific Nucleases: Potential Applications for Crop Improvement.

Humans are currently facing the problem of how to ensure that there is enough food to feed all of the world's population. Ensuring that the food supply is sufficient will likely require the modification of crop genomes to improve their agronomic traits. The development of engineered sequence-specific nucleases (SSNs) paved the way for targeted gene editing in organisms, including plants.

Origin of Jupiter's cloud-level zonal winds remains a puzzle even after Juno.

How far Jupiter's cloud-level zonal winds penetrate into its interior, a question related to the origin of the winds, has long been a major puzzle about Jupiter. There exist two different views: the shallow scenario in which the cloud-level winds are confined within the thin weather layer at cloud top and the deep scenario in which the cloud-level winds manifest thermal convection in the deep interior. We interpret, using two different models corresponding to the two scenarios, the high-precision measurements of Jupiter's equatorially antisymmetric gravitational field by the Juno spacecraft.

Using Jupiter's gravitational field to probe the Jovian convective dynamo.

Convective motion in the deep metallic hydrogen region of Jupiter is believed to generate its magnetic field, the strongest in the solar system. The amplitude, structure and depth of the convective motion are unknown. A promising way of probing the Jovian convective dynamo is to measure its effect on the external gravitational field, a task to be soon undertaken by the Juno spacecraft.

Precessing cylinders at the second and third resonance: turbulence controlled by geostrophic flow.

We investigate, via both asymptotic analysis and direct numerical simulation, precessionally driven flow of a homogeneous fluid confined in fluid-filled circular cylinders that rotate rapidly about their symmetry axis and precess about a different axis and that are marked by radius-height aspect ratios Γ=1.045945 and Γ=1.611089.

Swimming motion of rod-shaped magnetotactic bacteria: the effects of shape and growing magnetic moment.

We investigate the swimming motion of rod-shaped magnetotactic bacteria affiliated with the Nitrospirae phylum in a viscous liquid under the influence of an externally imposed, time-dependent magnetic field. By assuming that fluid motion driven by the translation and rotation of a swimming bacterium is of the Stokes type and that inertial effects of the motion are negligible, we derive a new system of the twelve coupled equations that govern both the motion and orientation of a swimming rod-shaped magnetotactic bacterium with a growing magnetic moment in the laboratory frame of reference. It is revealed that the initial pattern of swimming motion can be strongly affected by the rate of the growing magnetic moment.