Journal: bioRxiv

Article Title: Gene editing without a genome: generation and validation of F0 CRISPR mutants in gastropod mollusc Crepidula fornicata

doi: 10.1101/2025.11.26.690768

Figure Lengend Snippet: (a) 3D printed mould to create 150 µm wide channels in agarose (channels highlighted with white arrow heads) . (b) Imprinted agarose dish with one-celled C. fornicata embryos lined up in 150 µm channels, viewed under a dissecting microscope, plastic coverslip used to break the needle also in view. (bi) Schematized cross section of the imprinted microinjection dish with embryo in microinjection channel.

Article Snippet: C. fornicata full-length RNA Circular Consensus Sequences (CCS) were generated by Novogene (79,255 reads).

Techniques: Microscopy, Microinjection

Journal: bioRxiv

Article Title: Gene editing without a genome: generation and validation of F0 CRISPR mutants in gastropod mollusc Crepidula fornicata

doi: 10.1101/2025.11.26.690768

Figure Lengend Snippet: ( a-c ) Anterior views of organogenesis stage C. fornicata embryos injected with sgRNA20 for cf-pax6 , scale bar = 100 µm. All embryos are siblings. Yellow arrows indicate eyes. ( a ) embryo with two eyes (live image). ( b ) embryo with one eye (fixed specimen). ( c ) embryo with no eyes (live specimen). ( d ) Percentage of surviving embryos at the organogenesis stage with 0, 1, or 2 eyes. Total number of surviving embryos in each treatment are displayed below each bar.

Article Snippet: C. fornicata full-length RNA Circular Consensus Sequences (CCS) were generated by Novogene (79,255 reads).

Techniques: Injection

Journal: Research

Article Title: A Circular RNA Promotes Tumor Metastasis through Stabilizing MSI2 Protein in Pancreatic Ductal Adenocarcinoma

doi: 10.34133/research.0918

Figure Lengend Snippet: CircPRKD3 physically interacts with MSI2 in PDAC cells. (A) Coomassie blue staining of cytoplasmic proteins pulled down by circPRKD3 antisense (AS) or sense (S) DNA probes in PANC-1 cells. (B) Representative MS/MS spectrum identifying the MSI2 peptide from MS analysis of the excised band. (C) RNA pull-down assays showing specific association of MSI2 with circPRKD3, as confirmed by immunoblot analysis (top) and RT-PCR detection of circRNA (bottom). PABPC1 and GAPDH proteins serve as nonspecific controls. (D) In vitro circularized sense strand (S) circPRKD3 specifically enriches endogenous MSI2 proteins from PANC-1 cell lysates. Probe loading was verified by streptavidin-HRP. (E) RIP assays demonstrating enrichment of circPRKD3 by anti-MSI2 antibody compared to IgG control. MYC mRNA and TUBA1B mRNA serve as positive and negative controls, respectively. (F) Schematic diagrams of MSI2 RNA-binding domains and truncated mutants. (G and H) Flag-RIP experiments were performed in PANC-1 cells expressing MSI2 truncated mutants. Expression and IP efficiency of MSI2 mutants were evaluated by immunoblot analysis using anti-Flag antibody (G). CircPRKD3 enrichment by MSI2 truncated mutants was measured by RT-qPCR analysis (H). (I and J) Flag-RIP assays conducted in PANC-1 cells expressing either WT MSI2 or RRM1 mutants. Immunoblot analysis with anti-Flag antibody confirmed expression and IP efficiency of MSI2 mutants (I). RT-qPCR quantification revealed reduced circPRKD3 enrichment by K22A or R100A mutants compared to WT MSI2 (J). MYC mRNA serves as positive controls. (K) Transwell migration assays demonstrated that K22A and R100A mutants impaired the pro-migratory effects of MSI2 in both PANC-1 and CFPAC-1 cells. ** P < 0.01.

Article Snippet: RNA circularization was performed by incubating the annealed products with T4 DNA ligase (NEB) at 16 °C overnight.

Techniques: Staining, Tandem Mass Spectroscopy, Western Blot, Reverse Transcription Polymerase Chain Reaction, In Vitro, Control, RNA Binding Assay, Expressing, Quantitative RT-PCR, Migration