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Chen, Kai

Publications and source records attributed to Chen, Kai.

At least 19 records

Directed evolution expands CRISPR–Cas12a genome-editing capacity

CRISPR-Cas12a enzymes are versatile RNA-guided genome-editing tools with applications encompassing viral diagnosis, agriculture, and human therapeutics. However, their dependence on a 5'-TTTV-3' protospacer adjacent motif (PAM) next to DNA target sequences restricts Cas12a's gene targeting capability to only ∼1% of a typical genome. To mitigate this constraint, we used a bacterial-based directed evolution assay combined with rational engineering to identify variants of Lachnospiraceae bacterium Cas12a with expanded PAM recognition. The resulting Cas12a variants use a range of noncanonical PAMs while retaining recognition of the canonical 5'-TTTV-3' PAM. In particular, biochemical and cell-based assays show that the variant Flex-Cas12a utilizes 5'-NYHV-3' PAMs that expand DNA recognition sites to ∼25% of the human genome. With enhanced targeting versatility, Flex-Cas12a unlocks access to previously inaccessible genomic loci, providing new opportunities for both therapeutic and agricultural genome engineering.

Ma, Enbo↗

Chiral kinematic theory and converse vortical effects

Response theories in condensed matter typically describe the response of an electron fluid to external electromagnetic fields, while perturbations on neutral particles are often designed to mimic such fields. Here, we study the response of fermions to a space-time-dependent velocity field, thereby sidestepping the issue of gauge charge. First, we use a semiclassical chiral kinematic theory to obtain the local density of current and extract the orbital magnetization. The theory immediately predicts a "converse vortical effect," defined as an orbital magnetization driven by linear velocity. It receives contributions from magnetic moments on the Fermi surface and the Berry curvature of the occupied bands. Then, transcending semiclassics via a complementary Kubo formalism reveals that the uniform limit of a clean system receives only the Berry curvature contribution while other limits sense the Fermi surface magnetic moments too. We propose CoSi as a candidate material and suggest magnetometry of a sample under a thermal gradient to detect the effect. Overall, our study sheds light on the effects of a space-time-dependent velocity field on electron fluids and paves the way for exploring quantum materials using new probes and perturbations.

Chen, Kai↗

Lung and liver editing by lipid nanoparticle delivery of a stable CRISPR–Cas9 ribonucleoprotein

Lipid nanoparticle (LNP) delivery of clustered regularly interspaced short palindromic repeat (CRISPR) ribonucleoproteins (RNPs) could enable high-efficiency, low-toxicity and scalable in vivo genome editing if efficacious RNP–LNP complexes can be reliably produced. Here we engineer a thermostable Cas9 from Geobacillus stearothermophilus (GeoCas9) to generate iGeoCas9 variants capable of >100× more genome editing of cells and organs compared with the native GeoCas9 enzyme. Furthermore, iGeoCas9 RNP–LNP complexes edit a variety of cell types and induce homology-directed repair in cells receiving codelivered single-stranded DNA templates. Using tissue-selective LNP formulations, we observe genome-editing levels of 16-37% in the liver and lungs of reporter mice that receive single intravenous injections of iGeoCas9 RNP–LNPs. In addition, iGeoCas9 RNPs complexed to biodegradable LNPs edit the disease-causing SFTPC gene in lung tissue with 19% average efficiency, representing a major improvement over genome-editing levels observed previously using viral or nonviral delivery strategies. These results show that thermostable Cas9 RNP–LNP complexes can expand the therapeutic potential of genome editing.

59 BASIC BIOLOGICAL SCIENCES↗

In situ oxidation of reduced graphene oxide membranes by peracetic acid for dye desalination

Graphene oxide (GO) membranes with tunable interlayer spacings are of interest for dye removal from salty textile wastewater, and the membranes are often reduced to improve their stability, which inevitably lowers water permeance. Herein, we demonstrate that reduced GO (rGO) membranes can be facilely modified using peracetic acid (PAA) in situ to dramatically enhance water permeance while retaining dye rejection. Specifically, PAA-modified membranes (PrGO) are synthesized by vacuum-filtering hydrazine-reduced rGO nanosheets onto Nylon substrate and then exposing them to PAA solutions. The effects of the rGO layer thickness, PAA content, and PAA exposure time on the membrane chemistry, nanostructures, and salt/dye separation properties are thoroughly examined. For example, the PAA oxidation of a 100 nm-thick rGO membrane for 10 min increases water permeance by 180 %, from 35 to 93 Liter m −2 h −1 bar −1 , and decreases Na 2 SO 4 rejection from 10 % to 3.3 % while retaining the rejection of Congo red at ≈99.7 %. The PrGO membranes exhibit stable water permeance and >99 % dye rejection in multi-cycle tests in a crossflow system, surpassing state-of-the-art GO membranes and showcasing their potential for practical applications.

Dye desalination↗

Rapid DNA unwinding accelerates genome editing by engineered CRISPR-Cas9

Thermostable clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated (Cas9) enzymes could improve genome-editing efficiency and delivery due to extended protein lifetimes. However, initial experimentation demonstrated Geobacillus stearothermophilus Cas9 (GeoCas9) to be virtually inactive when used in cultured human cells. Laboratory-evolved variants of GeoCas9 overcome this natural limitation by acquiring mutations in the wedge (WED) domain that produce >100-fold-higher genome-editing levels. Cryoelectron microscopy (cryo-EM) structures of the wild-type and improved GeoCas9 (iGeoCas9) enzymes reveal extended contacts between the WED domain of iGeoCas9 and DNA substrates. Biochemical analysis shows that iGeoCas9 accelerates DNA unwinding to capture substrates under the magnesium-restricted conditions typical of mammalian but not bacterial cells. These findings enabled rational engineering of other Cas9 orthologs to enhance genome-editing levels, pointing to a general strategy for editing enzyme improvement. Together, these results uncover a new role for the Cas9 WED domain in DNA unwinding and demonstrate how accelerated target unwinding dramatically improves Cas9-induced genome-editing activity.

59 BASIC BIOLOGICAL SCIENCES↗

Author Correction: Engineering self-deliverable ribonucleoproteins for genome editing in the brain

Correction to: Nature Communicationshttps://doi.org/10.1038/s41467-024-45998-2, published online 26 February 2024. In the Acknowledgements section of this article, the grant number relating to National Institutes of Health funding to J.A.D. was incorrectly given as RM1HG009490 and should have been U19NS132303. The grant number 2334028 relating to the National Science Foundation funding to J.A.D. was omitted. Funding from Hampton University Summer Undergraduate Research Program, Mr. Li Ka Shing, Emerson Collective and the Innovative Genomics Institute (IGI) were omitted. The original article has been corrected.

60 APPLIED LIFE SCIENCES↗

Engineering self-deliverable ribonucleoproteins for genome editing in the brain

The delivery of CRISPR ribonucleoproteins (RNPs) for genome editing in vitro and in vivo has important advantages over other delivery methods, including reduced off-target and immunogenic effects. However, effective delivery of RNPs remains challenging in certain cell types due to low efficiency and cell toxicity. To address these issues, we engineer self-deliverable RNPs that can promote efficient cellular uptake and carry out robust genome editing without the need for helper materials or biomolecules. Screening of cell-penetrating peptides (CPPs) fused to CRISPR-Cas9 protein identifies potent constructs capable of efficient genome editing of neural progenitor cells. Further engineering of these fusion proteins establishes a C-terminal Cas9 fusion with three copies of A22p, a peptide derived from human semaphorin-3a, that exhibits substantially improved editing efficacy compared to other constructs. We find that self-deliverable Cas9 RNPs generate robust genome edits in clinically relevant genes when injected directly into the mouse striatum. Overall, self-deliverable Cas9 proteins provide a facile and effective platform for genome editing in vitro and in vivo.

59 BASIC BIOLOGICAL SCIENCES↗

A strong fracture-resistant high-entropy alloy with nano-bridged honeycomb microstructure intrinsically toughened by 3D-printing

Strengthening materials via conventional “top-down” processes generally involves restricting dislocation movement by precipitation or grain refinement, which invariably restricts the movement of dislocations away from, or towards, a crack tip, thereby severely compromising their fracture resistance. In the present study, a high-entropy alloy Al0.5CrCoFeNi is produced by the laser powder-bed fusion process, a “bottom-up” additive manufacturing process similar to how nature builds structures, with the microstructure resembling a nano-bridged honeycomb structure consisting of a face-centered cubic (fcc) matrix and an interwoven hexagonal net of an ordered body-centered cubic B2 phase. While the B2 phase, combined with high-dislocation density and solid-solution strengthening, provides strength to the material, the nano-bridges of dislocations connecting the fcc cells, i.e., the channels between the B2 phase on the cell boundaries, provide highways for dislocation movement away from the crack tip. Consequently, the nature-inspired microstructure imparts the material with an excellent combination of strength and toughness.

36 MATERIALS SCIENCE↗

Chalcogenide phase-change material advances programmable terahertz metamaterials: a non-volatile perspective for reconfigurable intelligent surfaces

Terahertz (THz) waves have gained considerable attention in the rising 6G communication due to their large bandwidth. However, the cost and power consumption become the major constraints for the commercialization of 6G THz systems as the frequency increases. Reconfigurable intelligent surface (RIS) comprising active metasurfaces and digital controllers has been proposed for beamforming in the 6G multiple-input-multiple-output systems, showing good potential to suppress the system size, weight, and power consumption (SWaP). Currently, their controlling diodes can hardly work up to THz frequencies. Therefore, several active stimuli have been investigated as alternatives. Among them, chalcogenide phase-change material Ge 2 Sb 2 Te 5 (GST) addresses large modulation depth, picosecond switching speed, and non-volatile properties. Notably, the non-volatile GST may enable RIS systems with memory and low control power. This work briefly reviews the advances of GST-tuned THz metamaterials (MTMs), discusses the current obstacles to overcome, and gives a perspective of GST applications in the rising 6G communications.

6G↗

Solid Electrolyte Bimodal Grain Structures for Improved Cycling Performance

Here, the application of solid-state electrolytes in Li batteries is hampered by the occurrence of Li-dendrite-caused short circuits. To avoid cell failure, the electrolytes can only be stressed with rather low current densities, severely restricting their performance. As grain size and pore distributions significantly affect dendrite growth in ceramic electrolytes such as Li 7 La 3 Zr 2 O 12 and its variants; here, a “detour and buffer” strategy to bring the superiority of both coarse and fine grains into play, is proposed. To validate the mechanism, a coarse/fine bimodal grain microstructure is obtained by seeding unpulverized large particles in the green body. The rearrangement of coarse grains and fine pores is fine-tuned by changing the ratio of pulverized and unpulverized powders. The optimized bimodal microstructure, obtained when the two powders are equally mixed, allows, without extra interface decoration, cycling for over 2000 h as the current density is increased from 1.0 mA·cm -2 , and gradually, up to 2.0 mA·cm -2 . The “detour and buffer” effects are confirmed from postmortem analysis. The complex grain boundaries formed by fine grains discourage the direct infiltration of Li. Simultaneously, the coarse grains further increase the tortuosity of the Li path. This study sheds light on the microstructure optimization for the polycrystalline solid-state electrolytes.

25 ENERGY STORAGE↗

High-bandwidth reconfigurable data acquisition card

A reconfigurable data acquisition card including at least one field programmable gate array (FPGA) and a configurable bus switch coupled with the FPGA. The bus switch forms at least first and second ports used by the FPGA, the bus switch being adaptable for insertion into a connection having a number of lanes at least equal to a combined number of lanes in the first and second ports. The data acquisition card further includes multiple optical transmitters and optical receivers. Each optical transmitter and optical receiver is coupled with a corresponding transceiver in the FPGA via at least one optical fiber having multiple communication links. Timing circuitry in the data acquisition card is coupled with clock generation and distribution circuitry in the FPGA and is configured to distribute clock and timing signals to detector front-ends with fixed latency and to synchronize input/output links with a system clock generated by the FPGA.

Chen, Kai↗

LaCeO x coupled N-doped graphene/Ru single-atoms as a binary-site catalyst for efficient hydrogen evolution based on hydrogen spillover

Based on the hydrogen spillover effect, versatile binary-site catalysts are promising for the alkaline hydrogen evolution reaction (HER). In which one site can adsorb and dissociate water, while the neighboring site is favorable for liberating hydrogen. Inspired by these possibilities, oxygen vacancy (O v )-rich LaCeO x coupled N-doped graphene/Ru single-atoms (LaCeO x @NGr/Ru 1 ) was synthesized as an efficient dual-site HER catalyst. It delivered an impressive low overpotential of 22 mV at a current density of 10mAcm –2 and a slight Tafel slope of 40 mV dec –1 in an alkaline medium, outstanding the advanced Ru-based catalysts. Moreover, this promising binary-component catalyst exhibited higher mass activity and longer lasting durability than commercial Pt/C catalyst (20 wt%). Finally, experimental and theoretical investigations provided insights into the HER mechanisms of LaCeO x @NGr/Ru 1 based on three indispensable steps: water adsorption and dissociation on O v -rich LaCeO x , diffusion of generated H* species towards Ce 3+ -N-Ru 1 bridges, and hydrogen evolution on Ru 1 sites.

36 MATERIALS SCIENCE↗

19.28% Efficiency and Stable Polymer Solar Cells Enabled by Introducing an NIR–Absorbing Guest Acceptor

Here, a near-infrared (NIR)-absorbing small-molecule acceptor (SMA) Y-SeNF with strong intermolecular interaction and crystallinity is developed by combining selenophene-fused core with naphthalene-containing end-group, and then as a custom-tailor guest acceptor is incorporated into the binary PM6:L8-BO host system. Y-SeNF shows a 65 nm red-shifted absorption compared to L8-BO. Thanks to the strong crystallinity and intermolecular interaction of Y-SeNF, the morphology of PM6:L8-BO:Y-SeNF can be precisely regulated by introducing Y-SeNF, achieving improved charge-transporting and suppressed non-radiative energy loss. Consequently, ternary polymer solar cells (PSCs) offer an impressive device efficiency of 19.28% with both high photovoltage (0.873 V) and photocurrent (27.88 mA cm –2 ), which is one of the highest efficiencies in reported single-junction PSCs. Notably, ternary PSC has excellent stability under maximum-power-point tracking for even over 200 h, which is better than its parental binary devices. The study provides a novel strategy to construct NIR-absorbing SMA for efficient and stable PSCs toward practical applications.

14 SOLAR ENERGY↗