Journal: MicrobiologyOpen

Article Title: Synergistic Integration of Recombinase Polymerase Amplification and CRISPR–Cas12a Enables High‐Sensitivity Detection of Mycoplasma bovis

doi: 10.1002/mbo3.70435

Figure Lengend Snippet: Optimization of RPA primers and CRISPR/Cas12a detection for the Mycoplasma bovis uvrC gene. (A) Schematic of p‐uvrC generation by restriction cloning of a 558‐bp uvrC fragment, which was used as the standard DNA template. (B) Screening of eight RPA primer pairs (PP1–PP8) targeting uvrC; PP3–PP8 produced strong, specific amplicons, and target amplification was also detected in DNA from M. bovis ‐positive bison lung. (C) Sequence of three CRISPR guide RNAs (gRNA1–gRNA3) designed within the amplified uvrC region adjacent to 5′‐TTTG‐3′ PAM sites. (D) Comparison of Cas12a fluorescence kinetics showed that all three guides detected uvrC, with gRNA3 producing the fastest and strongest signal. (E) Optimization of ssDNA‐FQ reporter concentration identified 0.75 μM as the best condition for visual detection with minimal background, although maximal fluorescence was reached at 5 μM. (F) All guide RNA combinations produced detectable fluorescence with similar kinetics, although gRNA2 + 3 and gRNA1 + 2 + 3 showed slightly faster onset.

Article Snippet: The target band was excised and purified using the Monarch DNA Gel Extraction Kit (New England Biolabs).

Techniques: CRISPR, Cloning, Screening, Produced, Amplification, Sequencing, Comparison, Fluorescence, Concentration Assay

Journal: iScience

Article Title: Tregs transduced with a Ro60-reactive TCR have increased immunosuppressive functions in Sjögren’s disease

doi: 10.1016/j.isci.2026.117642

Figure Lengend Snippet:

Article Snippet: DNA Plasmid Extraction Kit , Qiagen , Cat No. 27104.

Techniques: Isolation, Cell Isolation, DNA Extraction, Quantitation Assay, Plasmid Preparation, Extraction, Recombinant, Expressing, Software

Journal: iScience

Article Title: Tregs transduced with a Ro60-reactive TCR have increased immunosuppressive functions in Sjögren’s disease

doi: 10.1016/j.isci.2026.117642

Figure Lengend Snippet:

Article Snippet: Genomic DNA Extraction Kit , New England Biolabs , Cat No. T3010L.

Techniques: Isolation, Cell Isolation, DNA Extraction, Quantitation Assay, Plasmid Preparation, Extraction, Recombinant, Expressing, Software

Journal: iMeta

Article Title: Domesticated bacterial origin genes regulate homeostasis of the insect gut microbiome and empower adaptation

doi: 10.1002/imt2.70179

Figure Lengend Snippet: Signatures of genes suspected to be horizontally transferred from bacteria to leafhoppers. (A) Genome and transcriptome coverage of RdCDC9 ( Id09G000860 ). The read depths of genome resequencing and RNA sequencing data are log‐transformed and plotted along the focal genomic region, with numbers indicating the range of depths. Splice junctions in RdCDC9 inferred from the RNA sequencing reads are indicated by red lines. The two fragments used for PCR cloning of genomic DNA are represented as red segments. (B) Gene structures of the CDC genes in three leafhopper species. Green and pink rectangles represent the CDSs and UTRs, respectively, and the black lines represent introns. (C) Genome fragments cloned from Ne. cincticeps , Ne. nigropictus , and R. dorsalis genomic DNA. For the intron‐bearing genes RdCDC6 and RdCDC9 , two pairs of primers targeting the intron regions were used. Bar plots represent the expression levels of all CDCs detected by RT‐qPCR. (D) Identification of the RdCDC protein by immunoprecipitation‐mass spectrometry (IP‐MS). The R. dorsalis adult lysate was immunoprecipitated using IgG or an RdCDC9 antibody and analyzed by mass spectrometry to identify candidate proteins. (E) Hierarchical clustering results of CDC family members based on protein similarity. All genes from leafhoppers are clustered together and grouped with the CDCs from Firmicutes and Actinobacteria, suggesting that they may have originated from an ancient horizontal gene transfer event. The TCDB accession number and the common name or species of origin are provided for each protein used to construct the phylogeny. Predictions of the protein domains revealed that all leafhopper CDCs contained a region belonging to the thiol‐activated cytolysin superfamily (IPR036359). (F) Heatmap showing the average relative synonymous codon usage (RSCU) of leafhopper CDCs , bacterial CDCs ( PFO , LLO , and SLO ), and conserved single‐copy genes of the three leafhopper species. The gradient from dark blue to dark red indicates increasing RSCU values from low to high. LLO, listeriolysin O; PFO, perfringolysin O; SLO, streptolysin O.

Article Snippet: DNA was extracted using an Animal Genomic DNA Quick Extraction Kit for PCR Analysis (Beyotime, D0065S).

Techniques: Bacteria, RNA Sequencing, Transformation Assay, PCR Cloning, Clone Assay, Expressing, Quantitative RT-PCR, Immunoprecipitation, Mass Spectrometry, Protein-Protein interactions, Construct