Light‐Guided Genetic Scissors Based on Phosphorene Quantum Dot

Laser & Photonics Review, Journal Year: 2024, Volume and Issue: 18(11)

Published: July 10, 2024

Abstract Genetic engineering faces persistent challenges in achieving precise Deoxyribonucleic acid (DNA) cleavage, especially with the limitations associated current enzyme‐based methods, exemplified by issues CRISPR technologies. This study introduces a groundbreaking approach: utilizing reactive oxygen species (ROS) generated Multiphoton Absorption (MPA)‐excited Black Phosphorus Quantum Dots (BPQDs) under femto‐second laser irradiation. innovative method not only allows excitation lower energy but also enhances overall efficiency. The integration of complementary RNA sequences facilitates high‐efficiency, site‐selective DNA cleavage system, named “TADPOLE” (Targeted Precision Oriented Laser Excision). Beyond its precision targeting arbitrary using quantum dots, TADPOLE harnesses unique multiphoton absorption property BPQDs, enabling lower‐energy light sources suitable for vivo applications future. Moreover, approach integrates guiding and ultrafast technology to provide control over local ROS generation minimal heat production. guarantees site‐specific while mitigating risk damage non‐targeted sequences. In summary, this catalyzes advancements enzyme‐free technologies, transformative implications genetic engineering, biotechnology, medicine. holistic precision, versatility, endurance presented open new avenues targeted gene therapies related fields.

Language: Английский

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Size exclusion chromatography of biopharmaceutical products: From current practices for proteins to emerging trends for viral vectors, nucleic acids and lipid nanoparticles

Journal of Chromatography A, Journal Year: 2024, Volume and Issue: 1722, P. 464862 - 464862

Published: April 1, 2024

The 21st century has been particularly productive for the biopharmaceutical industry, with introduction of several classes innovative therapeutics, such as monoclonal antibodies and related compounds, gene therapy products, RNA-based modalities. All these new molecules are susceptible to aggregation fragmentation, which necessitates a size variant analysis their comprehensive characterization. Size exclusion chromatography (SEC) is one reference techniques that can be applied. analytical mAbs now well established some them emerging newer In this context, objective review article is: i) provide short historical background on SEC, ii) suggest clear guidelines selection packing material mobile phase successful method development in modern SEC; iii) highlight recent advances use narrow-bore micro-bore columns, ultra-wide pore low-adsorption column hardware. Some important innovations, recycling coupling SEC mass spectrometry, alternative detectors charge detection spectrometry photometry also described. addition, discusses multidimensional setups shows most at preparative scale. third part article, possibility characterization modalities reviewed. final summary opportunities limitations different products.

Language: Английский

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Rapid DNA unwinding accelerates genome editing by engineered CRISPR-Cas9

Cell, Journal Year: 2024, Volume and Issue: 187(13), P. 3249 - 3261.e14

Published: May 22, 2024

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

Language: Английский

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Unraveling host regulation of gut microbiota through the epigenome–microbiome axis

Trends in Microbiology, Journal Year: 2024, Volume and Issue: 32(12), P. 1229 - 1240

Published: June 5, 2024

Recent studies of dynamic interactions between epigenetic modifications a host organism and the composition or activity its associated gut microbiota suggest an opportunity for to shape microbiome through alterations that lead changes in gene expression noncoding RNA activity. We use insights from microbiota-induced review potential epigenetically regulate microbiome, which bidirectional 'epigenome–microbiome axis' emerges. This axis embeds environmentally induced variation, may influence adaptive evolution host–microbe interactions. furthermore present our perspective on how epigenome–microbiome can be understood investigated within holo-omic framework with applications applied health food sciences.

Language: Английский

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The pAblo·pCasso self-curing vector toolset for unconstrained cytidine and adenine base-editing in Gram-negative bacteria

Nucleic Acids Research, Journal Year: 2024, Volume and Issue: 52(4), P. e19 - e19

Published: Jan. 5, 2024

Abstract A synthetic biology toolkit, exploiting clustered regularly interspaced short palindromic repeats (CRISPR) and modified CRISPR-associated protein (Cas) base-editors, was developed for genome engineering in Gram-negative bacteria. Both a cytidine base-editor (CBE) an adenine (ABE) have been optimized precise single-nucleotide modification of plasmid targets. CBE comprises deaminase conjugated to Cas9 nickase from Streptococcus pyogenes (SpnCas9), resulting C→T (or G→A) substitutions. Conversely, ABE consists fused SpnCas9 A→G T→C) editing. Several nucleotide substitutions were achieved using these plasmid-borne base-editing systems novel protospacer adjacent motif (PAM)-relaxed (SpRY) variant. Base-editing validated Pseudomonas putida other bacteria by inserting premature STOP codons into target genes, thereby inactivating both fluorescent proteins metabolic (antibiotic-resistance) functions. The functional knockouts obtained via reverted the wild-type genotype ABE. Additionally, series induction-responsive vectors facilitate curing platform single cultivation step, simplifying complex strain programs without relying on homologous recombination yielding plasmid-free, bacterial cells.

Language: Английский

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CRISPR/Cas12a-based biosensors for environmental monitoring and diagnostics

Environmental Technology & Innovation, Journal Year: 2024, Volume and Issue: 34, P. 103625 - 103625

Published: April 4, 2024

Contaminants, such as nucleic acids or toxic small molecules, threaten both human health and ecosystems when they infiltrate the environment. The precise highly sensitive identification of contaminants holds paramount importance across diverse domains, including safeguarding food integrity, facilitating clinical diagnostics, monitoring environmental conditions. Traditional methodologies, encompassing spectroscopy, chromatography, sequencing, metagenomics, have conventionally served pivotal roles in detection processes. Nevertheless, these methods encountered recurring challenges related to sensitivity, specificity, portability. This review focuses on groundbreaking CRISPR/Cas12-based biosensors. These biosensors leverage incredible precision programmability CRISPR/Cas system recognize specific targets. Here, we comprehensively assess fundamental mechanisms that enable detection, ranging from guide RNA design collateral cleavage. versatility CRISPR/Cas12 becomes evident through their applications. applications encompass medical safety, monitoring. transition conventional ultimately represents a significant milestone contaminant detection. By incorporating molecular biology, nanotechnology, bioinformatics, potential reshape landscape water CRIPSR-Cas diagnostics is transformative technology paves way for safer healthier future environment life.

Language: Английский

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A roadmap for affordable genetic medicines

Nature, Journal Year: 2024, Volume and Issue: 634(8033), P. 307 - 314

Published: July 17, 2024

Language: Английский

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Engineering self-deliverable ribonucleoproteins for genome editing in the brain

Nature Communications, Journal Year: 2024, Volume and Issue: 15(1)

Published: Feb. 26, 2024

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

Language: Английский

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Precise fine-turning of GhTFL1 by base editing tools defines ideal cotton plant architecture

Genome biology, Journal Year: 2024, Volume and Issue: 25(1)

Published: Feb. 26, 2024

CRISPR/Cas-derived base editor enables precise editing of target sites and has been widely used for basic research crop genetic improvement. However, the efficiency editors at different targets varies greatly.

Language: Английский

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Revisiting growth–defence trade‐offs and breeding strategies in crops

Plant Biotechnology Journal, Journal Year: 2024, Volume and Issue: 22(5), P. 1198 - 1205

Published: Feb. 27, 2024

Plants have evolved a multi-layered immune system to fight off pathogens. However, activation is costly and often associated with growth development penalty. In crops, yield the main breeding target usually affected by high disease resistance. Therefore, proper balance between defence critical for achieving efficient crop improvement. This review highlights recent advances in attempts designed alleviate trade-offs resistance crops mediated (R) genes, susceptibility (S) genes pleiotropic genes. We also provide an update on strategies optimizing growth-defence breed future desirable yield.

Language: Английский

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