Journal: Discover Oncology

Article Title: CRISPR/Cas technologies in pancreatic cancer research and therapeutics: recent advances and future outlook

doi: 10.1007/s12672-025-03383-5

Figure Lengend Snippet: Generation of pancreatic cancer models employing a range of CRISPR systems. Various CRISPR gene editing techniques are instrumental in generating GEMMs. Among them, CRISPR/Cas9 plays a pivotal role in the creation of transplantation models, either by modifying the genome of pancreatic cancer cells or by manipulating the immune system to facilitate PDX models. This system has also been widely applied in generating transgenic pancreatic cancer cell lines and genetically modified organoids, advancing research in cancer biology and therapeutic development

Article Snippet: 2 μg/ml puromycin at different time point (Day 15, 27, 31, 35) , HPAF-II, AsPC-1, PaTu8988S , Knockout , Toronto KnockOut (TKO) CRISPR Library (addgene No. 1000000069) , Wnt pathway genes, FZD5 , CRISPR knockout, Antibody-mediated inhibition , -Reduced proliferation , [ ] .

Techniques: CRISPR, Transplantation Assay, Transgenic Assay, Genetically Modified

Journal: Discover Oncology

Article Title: CRISPR/Cas technologies in pancreatic cancer research and therapeutics: recent advances and future outlook

doi: 10.1007/s12672-025-03383-5

Figure Lengend Snippet: Pooled CRISPR screening approaches in different experimental conditions. In direct in vivo screening, CRISPR is delivered into living organisms (e.g., mice) to induce genetic modifications in their natural biological context. In indirect in vivo screening, CRISPR is applied to cell lines or organoids derived from the in vivo model, which are then reintroduced into the organism, allowing for controlled exploration of genetic modifications. In vitro CRISPR screening is conducted in cultured cells for high-throughput gene editing and analysis of specific genetic targets. Sequencing technologies, such as NGS, are then used to identify novel oncogenes and druggable targets, providing insights into gene functions and the impact of specific genetic changes in isolated cells

Article Snippet: 2 μg/ml puromycin at different time point (Day 15, 27, 31, 35) , HPAF-II, AsPC-1, PaTu8988S , Knockout , Toronto KnockOut (TKO) CRISPR Library (addgene No. 1000000069) , Wnt pathway genes, FZD5 , CRISPR knockout, Antibody-mediated inhibition , -Reduced proliferation , [ ] .

Techniques: CRISPR, In Vivo, Derivative Assay, In Vitro, Cell Culture, High Throughput Screening Assay, Sequencing, Isolation

Journal: Discover Oncology

Article Title: CRISPR/Cas technologies in pancreatic cancer research and therapeutics: recent advances and future outlook

doi: 10.1007/s12672-025-03383-5

Figure Lengend Snippet: Therapeutic applications of CRISPR-driven gene editing. Through the targeted modification of key factors involved in the pathogenesis of pancreatic cancer, CRISPR systems, employing diverse mechanisms, offer promising prospects for advancing therapeutic strategies in the treatment of pancreatic cancer. PC pancreatic cancer

Article Snippet: 2 μg/ml puromycin at different time point (Day 15, 27, 31, 35) , HPAF-II, AsPC-1, PaTu8988S , Knockout , Toronto KnockOut (TKO) CRISPR Library (addgene No. 1000000069) , Wnt pathway genes, FZD5 , CRISPR knockout, Antibody-mediated inhibition , -Reduced proliferation , [ ] .

Techniques: CRISPR, Modification

Journal: Molecular cell

Article Title: CRAMP1 drives linker histone expression to enable Polycomb repression.

doi: 10.1016/j.molcel.2025.05.031

Figure Lengend Snippet: Figure 1. A genome-wide CRISPR-Cas9 genetic screen identifies an essential requirement for CRAMP1 and histone H1.4 in PRC2-mediated reporter repression (A) Schematic representation of GFP reporter repression by the PRC2 complex. (B) The GFP reporter is derepressed upon CRISPR-Cas9-mediated gene disruption of any of the three core PRC2 subunits, as assayed by flow cytometry. (C) A genome-wide CRISPR-Cas9 screen to identify factors required for PRC2 function. Following Cas9 expression in KBM-7 cells harboring the PRC2-sensitive GFP reporter, genome-wide mutagenesis was carried out with the Sabatini/Lander single guide RNA (sgRNA) library, 36 and GFP + cells isolated through two sequential rounds of FACS. ‘‘Significance’’ on the y axis represents the negative log of the ‘‘pos|score’’ metric reported by Model-based Analysis of Genome-wide CRISPR-Cas9 Knockout (MAGeCK). 37

Article Snippet: Single guide RNA (sgRNA) sequences were selected from the Sabatini/Lander Human CRISPR Pooled Library (Addgene #1000000100, kindly deposited by David Sabatini and Eric Lander 81 ) or the Brunello Human CRISPR Knockout Pooled Library (Addgene #73178, kindly deposited by David Root and John Doench 82 ).

Techniques: Genome Wide, CRISPR, Disruption, Flow Cytometry, Expressing, Mutagenesis, Isolation, Knock-Out

Journal: Molecular cell

Article Title: CRAMP1 drives linker histone expression to enable Polycomb repression.

doi: 10.1016/j.molcel.2025.05.031

Figure Lengend Snippet: Figure 5. Linker histones are not enriched at regions marked by H3K9me3 (A–D) Lack of linker histone enrichment at H3K9me3-marked genomic regions. (A) Tornado plots depicting linker histone CUT&Tag signal across H3K9me3 peaks from the ENCODE project; average signal intensity is shown in (B). (C) Heatmap depicting the lack of correlation between linker histone occupancy and H3K9me3. Cells are annotated with pairwise Spearman correlation coefficients. An example locus is shown in (D). (E) CUT&Tag faithfully profiles H3K9me3. Example loci comparing CUT&Tag versus H3K9me3 ChIP-seq data (ENCODE) are shown. (F and G) Linker histone insufficiency does not impair H3K9me3-dependent LINE-1 silencing by the HUSH complex. (F) Schematic representation of the dual- color reporter cell line designed to monitor both H3K9me3-dependent repression by the HUSH complex and linker histone-mediated PRC2-reporter repression. (G) HUSH-mediated LINE-1 silencing is unaffected upon CRAMP1 depletion. The indicated CRISPR sgRNAs were expressed in the dual-color reporter cell line, and GFP and iRFP fluorescence assayed by flow cytometry. See also Figure S5 and Table S2.

Article Snippet: Single guide RNA (sgRNA) sequences were selected from the Sabatini/Lander Human CRISPR Pooled Library (Addgene #1000000100, kindly deposited by David Sabatini and Eric Lander 81 ) or the Brunello Human CRISPR Knockout Pooled Library (Addgene #73178, kindly deposited by David Root and John Doench 82 ).

Techniques: ChIP-sequencing, CRISPR, Fluorescence, Flow Cytometry