Journal: International Journal of Molecular Sciences

Article Title: Leptin-Upregulated Metastasis-Associated Protein 1 Promotes Vasculogenic Mimicry in Breast Cancer Cells

doi: 10.3390/ijms26125726

Figure Lengend Snippet: Leptin upregulates MTA1 expression in human breast cancer cells. MTA1 mRNA expression was analyzed using qPCR in MDA-MB-231 ( A ) and Hs 578T cells ( B ) after leptin treatment for 24 h. Results are expressed as mean ± SD; * p < 0.05 and ** p < 0.01 vs. untreated control. MTA1 protein expression was assessed using Western blot in MDA-MB-231 ( C ) and Hs 578T cells ( D ) after 24 h of leptin treatment.

Article Snippet: MDA-MB-231 cells (1.5 × 10 5 ) and Hs 578T cells (2.0 × 10 5 ) were cultured under optimal conditions and transfected with 1 μg of the control or 0.5 μg of the MTA1 CRISPR activation plasmid (Santa Cruz, Danvers, MA, USA) for 48 h. Transfection was performed using the UltraCruz transfection reagent (Santa Cruz), according to the manufacturer’s protocol.

Techniques: Expressing, Control, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Leptin-Upregulated Metastasis-Associated Protein 1 Promotes Vasculogenic Mimicry in Breast Cancer Cells

doi: 10.3390/ijms26125726

Figure Lengend Snippet: Leptin upregulates MTA1 expression via the Ob-R/STAT3 pathway in human breast cancer cells. MTA1 protein expression was evaluated using Western blot in MDA-MB-231 ( A , C ) and Hs 578T cells ( B , D ) after 24 h of leptin treatment in the presence or absence of Ob-R BP ( A , B ) or AG490 ( C , D ).

Article Snippet: MDA-MB-231 cells (1.5 × 10 5 ) and Hs 578T cells (2.0 × 10 5 ) were cultured under optimal conditions and transfected with 1 μg of the control or 0.5 μg of the MTA1 CRISPR activation plasmid (Santa Cruz, Danvers, MA, USA) for 48 h. Transfection was performed using the UltraCruz transfection reagent (Santa Cruz), according to the manufacturer’s protocol.

Techniques: Expressing, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Leptin-Upregulated Metastasis-Associated Protein 1 Promotes Vasculogenic Mimicry in Breast Cancer Cells

doi: 10.3390/ijms26125726

Figure Lengend Snippet: MTA1 overexpression promotes VM in human breast cancer cells. MDA-MB-231 ( A , C , E ) and Hs 578T cells ( B , D , F ) were transfected with the MTA1 CRISPR activation plasmid for 48 h. MTA1 protein expression was assessed using Western blot in MDA-MB-231 ( A ) and Hs 578T cells ( B ). VM was performed using a 3D culture assay for 16 h in MDA-MB-231 ( C ) and Hs 578T cells ( D ) (40× magnification; scale bar = 500 μm). Results are expressed as mean ± SD; *** p < 0.001 vs. control plasmid. Expression of VM-related proteins was evaluated using Western blot in MDA-MB-231 ( E ) and Hs 578T cells ( F ).

Article Snippet: MDA-MB-231 cells (1.5 × 10 5 ) and Hs 578T cells (2.0 × 10 5 ) were cultured under optimal conditions and transfected with 1 μg of the control or 0.5 μg of the MTA1 CRISPR activation plasmid (Santa Cruz, Danvers, MA, USA) for 48 h. Transfection was performed using the UltraCruz transfection reagent (Santa Cruz), according to the manufacturer’s protocol.

Techniques: Over Expression, Transfection, CRISPR, Activation Assay, Plasmid Preparation, Expressing, Western Blot, Control

Journal: International Journal of Molecular Sciences

Article Title: Leptin-Upregulated Metastasis-Associated Protein 1 Promotes Vasculogenic Mimicry in Breast Cancer Cells

doi: 10.3390/ijms26125726

Figure Lengend Snippet: MTA1 silencing inhibits leptin-induced VM in human breast cancer cells. MDA-MB-231 ( A , C , E ) and Hs 578T cells ( B , D , F ) were treated with leptin 48 h after transfection with MTA1 siRNA. MTA1 protein expression was evaluated using Western blot in MDA-MB-231 ( A ) and Hs 578T cells ( B ). VM was performed using a 3D culture assay for 16 h in MDA-MB-231 ( C ) and Hs 578T cells ( D ) (40× magnification; scale bar = 250 μm). Results are expressed as mean ± SD; *** p < 0.001 vs. untreated control; ### p < 0.001 vs. control siRNA. Expression of VM-related proteins was evaluated using Western blot in MDA-MB-231 ( E ) and Hs 578T cells ( F ).

Article Snippet: MDA-MB-231 cells (1.5 × 10 5 ) and Hs 578T cells (2.0 × 10 5 ) were cultured under optimal conditions and transfected with 1 μg of the control or 0.5 μg of the MTA1 CRISPR activation plasmid (Santa Cruz, Danvers, MA, USA) for 48 h. Transfection was performed using the UltraCruz transfection reagent (Santa Cruz), according to the manufacturer’s protocol.

Techniques: Transfection, Expressing, Western Blot, Control

Journal: bioRxiv

Article Title: Macropinocytosis mediates neurotropism of Cryptococcus neoformans in a human organoid model of the blood-brain barrier

doi: 10.1101/2025.09.23.678106

Figure Lengend Snippet: a Representative fluorescent image of BBB organoids formed from EphA2 knockout (KO) human brain endothelial cells. EphA2 KO cells express RFP (magenta) against DAPI (blue) counterstaining. b Western blot analysis (1:1000 dilution of rabbit anti-Human EphA2; Cell signaling technology Inc. D4A2) (1:10,000 dilution of Goat Anti-Rabbit IgG H&L HRPab6721; Abcam Inc., Cambridge, MA, USA) confirming loss of EphA2 protein expression in EphA2 KO organoids, in contrast to controls (EphA2+ and EphA2 over-expression (o/e)). c Internalization of Cn ( Cryptococcus neoformans ) by BBB organoids is dependent on EphA2 expression. Organoids (EphA2+ or EphA2 KO brain endothelial cells) were exposed to CFSE-stained Cn for 48 h, after which organoids were sectioned and analyzed for Cn invasion (Welch two-sample t-test: t = 4.28, df = 13.94, p < 0.0008)(R v.4.4.2). d Comparison of GFAP expression between BBB organoids formed with EphA2 KO versus EphA2+ brain endothelial cells. Organoids deficient in EphA2 show significantly higher levels of GFAP expression compared to EphA2+ organoids. Quantification was performed on GFAP-antibody probed organoid sections by applying a uniform threshold to images and measuring the percent area in the relevant channel above the threshold (Welch two-sample t-test: t = 2.42, df = 10.68, p = 0.0343)(R v.4.2.2). e Organoids exposed to Cd ( Cryptococcus deuterogattii ) show increased GFAP expression in stark contrast to organoids exposed to Cn after 48 h of exposure (Welch two-sample t-test comparing Cn & Cd treatments: t = 3.84, df = 9.38, p = 0.003, comparing Cd & Control treatments: t = 4.25, df = 10.57, p = 0.0015)(R v.4.4.2). For panels c – e, each data point represents the average value for an organoid across several images.

Article Snippet: Two sets of plasmids were used in the CRISPR/Cas9 method - the EphA2 CRISPR/Cas9 KO plasmids and the EphA2 HDR plasmids, both of which were ordered from Santa Cruz Biotechnology.

Techniques: Knock-Out, Western Blot, Expressing, Over Expression, Staining, Comparison, Control

Journal: bioRxiv

Article Title: Macropinocytosis mediates neurotropism of Cryptococcus neoformans in a human organoid model of the blood-brain barrier

doi: 10.1101/2025.09.23.678106

Figure Lengend Snippet: a Cn induced stimulation of GTP-bound Cdc42 is contingent on EphA2 expression and b CD44 engagement. EphA2+ and EphA2 KO human brain endothelial cells were treated with either a Cn or b cps1Δ Cn deletion mutant at an MOI of 10 for 30 min and then lysed. Untreated and treated cell lysates were analyzed for GTP bound Cdc42 (active Cdc42) using the Cytoskeleton G-LISA assay. Data are expressed as a mean ± SD of three independent experiments. Statistical analysis was done using a two tailed unpaired t-test (n = 8, t = 0.2156, df = 14, **** p < 0.0001)(GraphPad Prism 10 software). c, d Expression of Cdc42 protein in lysates was confirmed by western blot analysis of nitrocellulose membrane probed with primary antibody to amino acid 150-182 of Cdc42 protein. (1:800 dilution of mouse anti-human Cdc42; cytoskeleton Inc. ACD04) (1:10,000 dilution of Goat anti-Mouse IgG H&L HRP ab6789; Abcam Inc., Cambridge, MA, USA). e, f Cn recruits a CD44-EphA2 protein complex in brain endothelial cells, detected by Proximity Labeling Assays (PLA - DuoLink) of EphA2 and CD44 in e mouse primary brain endothelial cells and f human brain endothelial cells (EphA2+, iBMEC). The in-situ interaction was quantified as the area in an image above a universally applied threshold. Significantly higher fluorescence was observed in Cn (+) versus Cn (-) treatments in both mouse primary cells (t = 4.40, df = 43.13, p < 0.0001) and human cells (t = 3.63, df = 131.88, p = 0.0004). In mouse primary cells, there was significantly higher signal in the Cn (+) compared to the Cd (+) treatments (t = 4.08, df = 43.63, p < 0.0002)(R v.4.4.2). Mouse primary brain endothelial cells were exposed to Cd , Cn media or a no primary antibody control. g Representative fluorescent images of PLA in human brain endothelial cells (EphA2+, iBMECs). Top row: iBMECs exposed to Cn for 90 min. Middle row: control panel iBMECs not exposed to Cn . Bottom row: control treatment in which iBMECs were exposed to Cn but omitting primary antibodies (anti-CD44 and anti-EphA2) Left column: Nuclear stain. Middle column: Fluorescent puncta (PLA-red probe) are indicative of target proteins (EphA2 and CD44) in proximity (within 40 nm) to each other. Right column: merged images.

Article Snippet: Two sets of plasmids were used in the CRISPR/Cas9 method - the EphA2 CRISPR/Cas9 KO plasmids and the EphA2 HDR plasmids, both of which were ordered from Santa Cruz Biotechnology.

Techniques: Expressing, Mutagenesis, Two Tailed Test, Software, Western Blot, Membrane, Labeling, In Situ, Fluorescence, Control, Staining

Journal: bioRxiv

Article Title: Macropinocytosis mediates neurotropism of Cryptococcus neoformans in a human organoid model of the blood-brain barrier

doi: 10.1101/2025.09.23.678106

Figure Lengend Snippet: Alpha-Fold-Multimer structure prediction indicates reliable binding between EphA2 and CD44 . a Cartoon representation of the AlphaFold3-predicted model of the EphA2 structure. extracellular region in complex with CD44, highlighting two distinct putative binding interfaces: the ligand-binding domain (LBD) (ipTM = 0.13; pTM = 0.49) and the fibronectin domains (FN1-FN2) (ipTM = 0.11; pTM = 0.49). b-c Representative binding conformations of the LBD: CD44 and FN: CD44 complexes obtained from molecular dynamics simulations. Shown here are the major conformational clusters derived from all replica trajectories. EphA2 is depicted in green and CD44 in red cartoon representations. For clarity, only the principle interfacial hydrogen bonds are displayed.

Article Snippet: Two sets of plasmids were used in the CRISPR/Cas9 method - the EphA2 CRISPR/Cas9 KO plasmids and the EphA2 HDR plasmids, both of which were ordered from Santa Cruz Biotechnology.

Techniques: Binding Assay, Ligand Binding Assay, Derivative Assay

Journal: bioRxiv

Article Title: Macropinocytosis mediates neurotropism of Cryptococcus neoformans in a human organoid model of the blood-brain barrier

doi: 10.1101/2025.09.23.678106

Figure Lengend Snippet: Upon binding to CD44, Cn promotes the association of CD44 with the ligand binding domain (LBD) and interactions involving both FN1 and FN2 domains, suggesting that CD44 can also engage EphA2 via a membrane-proximal site and/or the fibronectin 2 domain (FN2) (black arrows). The CD44-EphA2 molecular interaction leads to a PKC-dependent phosphorylation of EphA2, and the downstream activation of GTP-bound Cdc42 (green arrows). Active Cdc42 stimulates membrane ruffling by remodeling the cytoskeleton via actin polymerization. The ensuing macropinocytosis of Cn is a result of macropinosomes large enough to internalize and transport Cn across the brain endothelium. Cd does not cross the brain endothelium (crossed-out black arrow), but stimulates the expression of GFAP+ astrocytes by as yet an unknown mechanism (dashed arrow). Figure created in BioRender.com.

Article Snippet: Two sets of plasmids were used in the CRISPR/Cas9 method - the EphA2 CRISPR/Cas9 KO plasmids and the EphA2 HDR plasmids, both of which were ordered from Santa Cruz Biotechnology.

Techniques: Binding Assay, Ligand Binding Assay, Membrane, Phospho-proteomics, Activation Assay, Expressing

Journal: iScience

Article Title: AKG-TET axis is central to senescence plasticity

doi: 10.1016/j.isci.2025.114298

Figure Lengend Snippet: AKG-TET deficiency leads to replicative senescence aHDFs (3 × 10 5 cells/mL) were seeded in each culture plate and were treated, in triplicate, without (UT) or treated (T) with C35 (5 μM), peptides (20 μM), TET1 siRNA ( TET1 i ; 300 nM). Cells treated with Bleomycin (Bleo; 5 μg/mL), or H 2 O 2 (100 μM) were used as positive control. Cells were incubated with 30 μM BrdU for 18 h prior to removal of both the cells and culture media following 7 days of treatment. (A) Heatmap of TET gene expression. (B) TET activity and global DNA levels of 5 mC, 5hmC, and 5 fC. (C) Expression patterns of energy/stress and nutrient-sensing pathways in aging PBMCs mirror those observed in young, actively replicating aHDFs (week 3) undergoing senescence, as well as those treated with TET1 i , C35, and RLS. (D) Expression level of hTERT , NAMPT , and PCNA . (E) Expression level of COL1A1 and ELN. (F) Cellular UPS activity, ATP, NAD + /NADH ratio, and LC3B levels. (G) Cellular levels of LC3B without (−) and after pre-treatment with (+) Bafilomycin A1 (1 μM) for 24 h. (H) Cellular ROS. (I) Extent of oxidative DNA damage assessed by the cellular level of 8OHdG. Positive control was comprised of cells treated with H 2 O 2 and Bleomycin (Bleo). Negative control was comprised of cells treated with CLV. (J) γH2AX immunolocalized to nuclei. For positive control, cells were treated with Bleomycin (Bleo). Intranuclear γH2AX (green) appears as focal spots (red arrows) or as a diffuse pan-nuclear pattern (yellow arrows) in nuclei marked by DAPI (blue) staining. Nuclei are further highlighted by double hashed lines. The boxes in the left pane (scale bars 10 μM) are magnified in the right insets (scale bars 2.5 μm). (K) NFKB1 gene expression. (L) Level of intra-nuclear phosphorylated p65 (p65P). (M) Cellular levels of ROS, IL6, and IL8 were secreted into culture media. (N) Expression levels of senescence markers ( CDKN2A , CDKN1A , CDKN1B , LTA4H , TIMP1 , and MMP1 ). (O) Expression level of LDH toxicity, SAβ-Gal activity, and extracellular level of lactic acid (LA). (P) LDHA1 gene expression. (Q) Rate of proliferation assessed by the quantitation of BrdU incorporated into the nuclei of cells in S-phase. Bar and line graphs show the means ± SD. Boxplots show the first and third quartiles and median values. Points are shown as empty and mean points as filled circles. The distribution of data points is shown by Beeswarm in Violin plots. Statistical significance was assessed using Student’s t test for two-group comparisons. p -values are presented as follows: ns (not significant), p ≤ 5 × 10 −1 , ∗p ≤ 5 × 10 −2 , ∗∗p ≤ 5 × 10 −3 , ∗∗∗p ≤ 5 × 10 −4 .

Article Snippet: CRISPR activation system for TET1 ( TET1 CR ) , Santa Cruz Biotechnology (Santa Cruz, CA) , sc-400845-ACT-2.

Techniques: Positive Control, Incubation, Gene Expression, Activity Assay, Expressing, Negative Control, Staining, Quantitation Assay

Journal: iScience

Article Title: AKG-TET axis is central to senescence plasticity

doi: 10.1016/j.isci.2025.114298

Figure Lengend Snippet: AKG-TET dependent resilience to damage and protection against damage-induced senescence Proliferating aHDFs (0.3 × 10 6 /mL) and PBMCs (70 years; PBMC 70Yr, 1 × 10 6 /mL) were seeded in culture plates and pre-treated in triplicate without (untreated: UT) or with H 2 O 2 (100 μM) for 24 h. After 24 h, cells were washed and treated without or with CLV (20 μM), or CRISPR TET1 ( TET1 CR ;1 μg/mL). Cells were incubated with 30 μM BrdU for 18 h prior to removal of both the cells and culture media following 7 days of treatment. (A) Expression levels of TETs in PBMCs. (B) AKG bioavailability in PBMCs. (C) Expression levels of energy/stress and nutritional sensors in PBMCs. (D) Heatmap of senescence marker gene expression in PBMCs. (E) Level of extracellular lactic acid (LA) and SAβ-Gal activity in PBMCs. (F) Quantification of BrdU and ROS (qROS) in PBMCs. Bar and line graphs show the means ± SD. Statistical significance was assessed using Student’s t test for two-group comparisons. p -values are presented as follows: ns (not significant), p ≤ 5 × 10 −1 , ∗p ≤ 5 × 10 −2 , ∗∗p ≤ 5 × 10 −3 , ∗∗∗p ≤ 5 × 10 −4 . Points are shown as empty and mean points as filled circles.

Article Snippet: CRISPR activation system for TET1 ( TET1 CR ) , Santa Cruz Biotechnology (Santa Cruz, CA) , sc-400845-ACT-2.

Techniques: CRISPR, Incubation, Expressing, Marker, Gene Expression, Activity Assay

Journal: iScience

Article Title: AKG-TET axis is central to senescence plasticity

doi: 10.1016/j.isci.2025.114298

Figure Lengend Snippet: AKG-TET deficient senescence state is reversible Young replicatively proliferating aHDFs (week 3, 0.3 × 10 6 /mL)) were seeded in culture plates and pre-treated in triplicates without (UT) and with C35 (5 μM), RLS (20 μM), or TET1 siRNA ( TET1 i ; 300 nM). Aliquot of cells were incubated with 30 μM BrdU for 18 h prior to removal of both the cells and culture media following 7 days of treatment. The remaining cells were re-seeded in equal numbers and cultured without treatment for an additional 7 days (withdrawal). Cultures were incubated with 30 μM BrdU for 18 h prior to removal of both the cells and culture media. (A) Quantification of TET activity, and 5 mC and 5fc levels. (B) Expression level of energy/stress and nutritional sensor genes. (C) Expression level of NFKB1 , RELA , IKBA , COL1A1, and ELN . (D) γH2AX immunolocalized (cyan) in nuclei. Nuclei stained for DAPI (deep blue) are marked by double hashed lines. (E) Extracellular level of lactic acid (LA), SAβ-Gal activity, LDH toxicity, and UPS. (F) Intracellular level of ROS and 8OHdG. (G) Heatmap of senescence markers and RRM2 gene expression. (H) Rate of proliferation assessed by BrdU incorporation into the nuclei of cells in S-phase. Bar and line graphs show the means ± SD. Boxplots show the first and third quartiles and median values. All points are shown as empty, and mean points as filled circles. The distribution of all data points is shown by Beeswarm in Violin plots. Statistical significance was assessed using Student’s t test for two-group comparisons. p -values are presented as follows: ns (not significant), p ≤ 5 × 10 −1 , ∗p ≤ 5 × 10 −2 , ∗∗p ≤ 5 × 10 −3 , ∗∗∗p ≤ 5 × 10 −4 .

Article Snippet: CRISPR activation system for TET1 ( TET1 CR ) , Santa Cruz Biotechnology (Santa Cruz, CA) , sc-400845-ACT-2.

Techniques: Incubation, Cell Culture, Activity Assay, Expressing, Staining, Gene Expression, BrdU Incorporation Assay

Journal: iScience

Article Title: AKG-TET axis is central to senescence plasticity

doi: 10.1016/j.isci.2025.114298

Figure Lengend Snippet: Activation of the AKG-TET axis reverses replicative and age induced senescence Replicatively induced senescence. Replicatively senescent aHDFs (0.3 × 10 6 cells/mL) were seeded in triplicate in each plate and were treated without (UT) and with RLS (20 μM), CLV (20 μM), or CRISPR TET1 plasmids (TET1 CR , 1 μg/mL). Cells were incubated with 30 μM BrdU for 18 h prior to removal of both the cells and culture media following 7 days of treatment. (A) Expression level of TET s. (B) AKG bioavailability and TET activity (TET act ). (C) Expression level of energy/stress and nutritional sensor gene expression. (D) Intracellular level of ROS. (E) Heatmap of senescence marker and RRM2 gene expression. (F) Level of lactic acid (LA) in culture media. (G) Cellular SAβ-Gal activity. (H) Quantitation of nuclear BrdU. (I) Total cell numbers. Age induced senescence. PBMCs (PBMC 70Yr ) were seeded (1 × 10 6 cells/mL) in triplicate in each well of a 24-well plate and were treated without (UT) and with RLS (20 μM), CLV (20 μM), or CRISPR TET1 plasmids ( TET1 CR : 1 μg/mL). Cells and culture media were removed on day 7 of treatment for analysis. (J) Quantitation of bioavailable AKG and TET activity in cells treated with RLS versus CLV. (K) qPCR quantitation of the expression level of TET s. (L) qPCR quantitation of the expression level of energy/stress and nutritional sensor gene expression. (M) Quantitation of BrdU incorporated into nuclei of cells in S-phase, lactic (LA) acid in culture media, cellular SAβ-Gal activity, and ROS. (N) Heatmap of senescence markers and RRM2 in PBMCs. Bar and line graphs show the means ± SD. Boxplots show the first and third quartiles and median values. All points are shown as empty, and mean points as filled circles. The distribution of all data points is shown by Beeswarm in Violin plots. Statistical significance was assessed using Student’s t test for two-group comparisons. p -values are presented as follows: ns (not significant), p ≤ 5 × 10 −1 , ∗p ≤ 5 × 10 −2 , ∗∗p ≤ 5 × 10 −3 , ∗∗∗p ≤ 5 × 10 −4 , ∗∗∗∗p ≤ 5 × 10 −5 .

Article Snippet: CRISPR activation system for TET1 ( TET1 CR ) , Santa Cruz Biotechnology (Santa Cruz, CA) , sc-400845-ACT-2.

Techniques: Activation Assay, CRISPR, Incubation, Expressing, Activity Assay, Gene Expression, Marker, Quantitation Assay

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