Journal: bioRxiv

Article Title: Elucidating genes sufficient for viral entry into cells through sequential genome-wide CRISPR activation screens

doi: 10.64898/2026.03.06.710083

Figure Lengend Snippet: A) NGFR was expressed in Jurkat C6 cells using CRISPRa, or alternatively, Jurkat cells using cDNA expression. NGFR -expressing or control cells were then inoculated with rabies pseudovirus encoding mCD19t. Flow cytometry was then performed to determine the percentage of infected cells. This revealed that NGFR expression significantly increased the susceptibility of Jurkat cells to rabies pseudovirus infection. B) L-SIGN or DC-SIGN were expressed in Jurkat or primary human CD4 + T cells using cDNA expression. L-SIGN -expressing, DC-SIGN -expressing, or control cells were inoculated with rabies pseudovirus encoding hEGFRt (a truncated mutant of human EGFR). Flow cytometry was then performed to determine the percentage of infected cells. This revealed that L-SIGN or DC-SIGN expression significantly increased the susceptibility of Jurkat cells and primary T cells to Ebola pseudovirus infection. C) L-SIGN or DC-SIGN were expressed in primary human CD4 + T cells using cDNA expression, and then L-SIGN -expressing, DC-SIGN -expressing, or control cells were inoculated with GFP -expressing Ebola virus or zsGreen -expressing Sudan virus under BSL4 containment. Flow cytometry was then performed to determine the percentage of infected cells. This revealed that L-SIGN or DC-SIGN expression significantly increased the susceptibility of primary T cells to authentic Ebola and Sudan virus infection.

Article Snippet: Cells were infected with pseudotyped lentiviruses, and 72 hours later, were harvested and stained with antibodies against the relevant surface marker: mouse CD19 (Miltenyi Biotec, 130-111-884), mouse H2Kk (Miltenyi Biotec, 130-117-235), or human EGFR (R&D Systems, FAB9577R-100), and viability staining was performed using DAPI (Thermo Fisher Scientific, D1306).

Techniques: Expressing, Control, Flow Cytometry, Infection, Mutagenesis, Virus

Journal: bioRxiv

Article Title: Elucidating genes sufficient for viral entry into cells through sequential genome-wide CRISPR activation screens

doi: 10.64898/2026.03.06.710083

Figure Lengend Snippet: A) NGFR was expressed in Jurkat C6 cells using CRISPRa, or alternatively, Jurkat cells using cDNA expression. NGFR -expressing or control cells were then inoculated with rabies pseudovirus encoding mCD19t. Flow cytometry was then performed to determine the percentage of infected cells. This revealed that NGFR expression significantly increased the susceptibility of Jurkat cells to rabies pseudovirus infection. As positive controls, flow cytometry was used to confirm successful delivery of the sgRNA construct as part of the CRISPRa workflow (as denoted by BFP expression) and that NGFR was expressed (upon cDNA expression). B) L-SIGN or DC-SIGN were expressed in primary human CD4 + T cells using cDNA expression. L-SIGN -expressing, DC-SIGN -expressing, or control cells were inoculated with rabies pseudovirus encoding hEGFRt (a truncated mutant of human EGFR). Flow cytometry was then performed to determine the percentage of infected cells. This revealed that L-SIGN or DC-SIGN expression significantly increased the susceptibility of Jurkat cells and primary T cells to Ebola pseudovirus infection. Cells expressing the highest levels of L-SIGN and DC-SIGN were preferentially infected by Ebola pseudovirus. C) L-SIGN or DC-SIGN were expressed in primary human CD4 + T cells using cDNA expression, and then L-SIGN -expressing, DC-SIGN -expressing, or control cells were inoculated with GFP -expressing Ebola virus or zsGreen -expressing Sudan virus under BSL4 containment. On days 0, 1, and 2 post-infection, flow cytometry was performed to determine the percentage of infected cells and qPCR was performed on cell culture supernatants to quantify viral genome replication. This revealed that L-SIGN or DC-SIGN expression enabled authentic Ebola and Sudan virus entry into primary human T cells, but viral genome replication was impaired, perhaps reflective of cell-intrinsic restriction factors.

Article Snippet: Cells were infected with pseudotyped lentiviruses, and 72 hours later, were harvested and stained with antibodies against the relevant surface marker: mouse CD19 (Miltenyi Biotec, 130-111-884), mouse H2Kk (Miltenyi Biotec, 130-117-235), or human EGFR (R&D Systems, FAB9577R-100), and viability staining was performed using DAPI (Thermo Fisher Scientific, D1306).

Techniques: Expressing, Control, Flow Cytometry, Infection, Construct, Mutagenesis, Virus, Cell Culture

Journal: Science Advances

Article Title: Discovery and preclinical evaluation of monoclonal antibodies and bispecific engagers targeting the NKG2A inhibitory receptor

doi: 10.1126/sciadv.adu0690

Figure Lengend Snippet: ( A ) Cell surface HLA-E expression level of A549, H2030, and 293T cells (left). ADCC activity of anti-EGFR hIgG1 cetuximab (ctx) with mona-IgG, 1B2 IgG, or 1B2-6 IgG in NSCLC A549, H2030 cells, and 293T cells (right graphs). ( B ) ADCC activity of anti-HER2 IgG1 pertuzumab with 1B2-6 IgG in NSCLC A549, H2030 cells, and 293T cells. (A and B) Lactate dehydrogenase (LDH) release assay results in presence of primary NK cells from healthy donor PBMCs. E:T ratio, 5:1. Significance was determined by unpaired two-tailed Student’s t test. * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001 versus ctx or pertuzumab (ptz) single treatment in vitro. (A) Cell surface HLA-E expression level of A549, H2030, and 293 T cells (left). ADCC activity of anti-EGFR hIgG1 cetuximab (ctx IgG1) with mona-IgG, 1B2 IgG, or 1B2-6 IgG in NSCLC A549, H2030 cells, and 293T cells (right graphs). (B) ADCC activity of anti-HER2 IgG1 pertuzumab (ptz IgG1) with 1B2-6 IgG in NSCLC A549, H2030 cells, and 293T cells. (A and B) LDH release assay results in presence of primary NK cells from healthy donor PBMCs. E:T ratio of 5:1. Significance was determined by unpaired two-tailed Student’s t test. * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001 versus ctx or ptz single treatment.

Article Snippet: Durvalumab (anti–PD-L1 antibody, A2013), cetuximab (anti-EGFR antibody, A2000), and pertuzumab (anti-HER2 antibody, A2008) were purchased from Selleck Chemicals.

Techniques: Expressing, Activity Assay, Lactate Dehydrogenase Assay, Two Tailed Test, In Vitro