One-step multiple gene disruption in the model organism Saccharomyces cerevisiae is a highly useful tool for both basic and applied research, but it remains a challenge. Here, we report a rapid, efficient, and potentially scalable strategy based on the type II Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-CRISPR associated proteins (Cas) system to generate multiple gene disruptions simultaneously in S. cerevisiae. A 100 bp dsDNA mutagenizing homologous recombination donor is inserted between two direct repeats for each target gene in a CRISPR array consisting of multiple donor and guide sequence pairs. An ultrahigh copy number plasmid carrying iCas9, a variant of wild-type Cas9, trans-encoded RNA (tracrRNA), and a homology-integrated crRNA cassette is designed to greatly increase the gene disruption efficiency. As proof of concept, three genes, CAN1, ADE2, and LYP1, were simultaneously disrupted in 4 days with an efficiency ranging from 27 to 87%. Another three genes involved in an artificial hydrocortisone biosynthetic pathway, ATF2, GCY1, and YPR1, were simultaneously disrupted in 6 days with 100% efficiency. This homology-integrated CRISPR (HI-CRISPR) strategy represents a powerful tool for creating yeast strains with multiple gene knockouts.

A range of novel two-dimensional materials have been actively explored for More Moore and More-than-Moore device applications because of their ability to form van der Waals heterostructures with unique electronic properties. However, most of the reported electronic devices exhibit insufficient control of multifunctional operations. Here, we leverage the band-structure alignment properties of narrow-bandgap black phosphorus and large-bandgap molybdenum disulfide to realize vertical heterostructures with an ultrahigh rectifying ratio approaching 10 and on-off ratio up to 10. Furthermore, we design and fabricate tunable multivalue inverters, in which the output logic state and window of the mid-logic can be controlled by specific pairs of channel length and, most importantly, by the electric field, which shifts the band-structure alignment across the heterojunction. Finally, high gains over 150 are achieved in the inverters with optimized device geometries, showing great potential for future logic applications.

SUMMARY Mitochondrial diseases include a group of maternally inherited genetic disorders caused by mutations in mitochondrial DNA (mtDNA). In most of these patients, mutated mtDNA coexists with wild type mtDNA, a situation known as mtDNA heteroplasmy. Here we report on a strategy towards preventing germline transmission of mitochondrial diseases by inducing mtDNA heteroplasmy shift through the selective elimination of mutated mtDNA. As a proof of concept we took advantage of NZB/BALB heteroplasmic mice, which contain two mtDNA haplotypes, BALB and NZB, and selectively prevented their germline transmission using either mitochondria targeted restriction endonucleases or TALENs. In addition, we successfully reduced human mutated mtDNA levels responsible for Leber’s hereditary optic neuropathy (LHOND), and neurogenic muscle weakness, ataxia and retinitis pigmentosa (NARP), in mammalian oocytes using mito-TALENs. Altogether, our approaches represent a potential therapeutic avenue for preventing the transgenerational transmission of human mitochondrial diseases caused by mutations in mtDNA.

The CCCTC-binding factor (CTCF) works together with the cohesin complex to drives the formation of chromatin loops and topologically associating domains, but its role in gene regulation has not been fully defined. Here, we investigated the effects of acute CTCF loss on chromatin architecture and transcriptional programs in mouse embryonic stem cells undergoing differentiation to neural precursor cells. We identified CTCF-dependent enhancer-promoter contacts genome-wide and found that they disproportionally affect genes that are bound by CTCF at promoter and dependent on long-distance enhancers. Disruption of promoter-proximal CTCF binding reduced both long-range enhancer-promoter contacts and transcription, which are restored by artificial tethering of CTCF to the promoter. Promoter-proximal CTCF binding is correlated with transcription of over 2,000 genes, across a diverse set of adult tissues. Taken together, our study shows that CTCF binding to promoters may promote long-distance-enhancer dependent transcription at specific genes in diverse cell types.

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On 4 and 6 July 2019, two large strike‐slip earthquakes with W‐phase moment magnitudes MWW 6.5 (foreshock) and MWW 7.1 (mainshock) struck the Eastern California Shear Zone, northeast of Ridgecrest. The faulting geometry and kinematic coseismic slip distribution of both events are determined by jointly inverting seismological and geodetic observations guided by aftershock and surface rupture locations. The foreshock ruptured two orthogonal faults with a prominent L‐shaped geometry with maximum slip of ~1.1 m on the NE‐SW segment. The mainshock faulting extended NW‐SE along several primary fault segments that straddle the foreshock slip. The surface rupture and slip model indicate mostly near‐horizontal strike‐slip motion with maximum slip of ~3.7 m, but there is a localized vertical dip‐slip motion. Both the foreshock and mainshock ruptures terminate in regions of complex surface offsets. High aftershock productivity and low rupture velocity may be the result of rupture of a relatively immature fault system.

Microplastic pollution in inland waters is receiving growing attentions. Reservoirs are suspected to be particularly vulnerable to microplastic pollution. However, very limited information is currently available on pollution characteristics of microplastics in reservoir ecosystems. This work studied the distribution and characteristics of microplastics in the backwater area of Xiangxi River, a typical tributary of the Three Gorges Reservoir. Microplastics were detected in both surface water and sediment with concentrations ranging from 0.55 × 10 to 342 × 10 items km and 80 to 864 items m, respectively. Polyethylene, polypropylene, and polystyrene were identified in surface water, whereas polyethylene, polypropylene, and polyethylene terephthalate, and pigments were observed in sediment. In addition, microplastics were also detected in the digestion tracts of 25.7% of fish samples, and polyethylene and nylon were identified. Redundancy analysis indicates a weak correlation between microplastics and water quality variables but a negative correlation with water level of the reservoir and Secchi depth. Results from this study confirm the presence of high abundance microplastics in reservoir impacted tributaries, and suggest that water level regulated hydrodynamic condition and input of nonpoint sources are important regulators for microplastic accumulation and distribution in the backwater area of reservoir tributaries.