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Promega haloprotac3

a Schematics illustrate five degron technologies for ligand-inducible protein depletion. b , c Western blots indicate comparative depletion of endogenously tagged CTCF and RAD21 proteins in KOLF2.2J iPSCs. The endogenous genes had a homozygous knock-in of mAID, miniIAA7, FKBP12(F36V), HaloTag, and IKZF3 degron tags at the C-termini of the target genes, and the target proteins were induced to degrade by treating cells with 1 μM 5-Phenyl-indole-3-acetic acid (5-Ph-IAA, synthetic auxin), 500 μM Indole-3-acetic acid (IAA, auxin), 1 μM dTAG13, 1 μM <t>HaloPROTAC3,</t> or 1 μM Pomalidomide, respectively, for 0, 1, 6, and 24 h. d Western blots show relative recovery rates of endogenous CTCF protein levels in five degron systems. KOLF2.2J iPSCs were treated with respective ligands for 6 h, then the ligands were washed out, and target protein levels were assessed after 24 h and 48 h with western blot. e Bar plots show quantification of target protein depletion after 1 h, 6 h, and 24 h of treatment with each chemical degrader. f The bar plot shows relative CTCF protein recovery after ligand washout in each degron system. g The single cell analysis (UMAP visualization) shows the relative expression levels of endogenous VHL , CRBN genes, and CAG-promoter driven OsTIR1 gene expression in iPSCs differentiated as embryoid bodies for 21 days. Each experiment in b – d was repeated independently at least twice with similar results. Data and error bars in e and f indicate the means ± SD ( n = 3). Source data is provided as a Source Data file.

Haloprotac3, supplied by Promega, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/haloprotac3/haloprotac3/pmc12274581-271-26-27
Average 90 stars, based on 1 article reviews

haloprotac3 - by Bioz Stars, 2026-09

90/100 stars

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1) Product Images from "Systematic comparison and base-editing-mediated directed protein evolution and functional screening yield superior auxin-inducible degron technology"

Article Title: Systematic comparison and base-editing-mediated directed protein evolution and functional screening yield superior auxin-inducible degron technology

Journal: Nature Communications

doi: 10.1038/s41467-025-61848-1

Figure Legend Snippet: a Schematics illustrate five degron technologies for ligand-inducible protein depletion. b , c Western blots indicate comparative depletion of endogenously tagged CTCF and RAD21 proteins in KOLF2.2J iPSCs. The endogenous genes had a homozygous knock-in of mAID, miniIAA7, FKBP12(F36V), HaloTag, and IKZF3 degron tags at the C-termini of the target genes, and the target proteins were induced to degrade by treating cells with 1 μM 5-Phenyl-indole-3-acetic acid (5-Ph-IAA, synthetic auxin), 500 μM Indole-3-acetic acid (IAA, auxin), 1 μM dTAG13, 1 μM HaloPROTAC3, or 1 μM Pomalidomide, respectively, for 0, 1, 6, and 24 h. d Western blots show relative recovery rates of endogenous CTCF protein levels in five degron systems. KOLF2.2J iPSCs were treated with respective ligands for 6 h, then the ligands were washed out, and target protein levels were assessed after 24 h and 48 h with western blot. e Bar plots show quantification of target protein depletion after 1 h, 6 h, and 24 h of treatment with each chemical degrader. f The bar plot shows relative CTCF protein recovery after ligand washout in each degron system. g The single cell analysis (UMAP visualization) shows the relative expression levels of endogenous VHL , CRBN genes, and CAG-promoter driven OsTIR1 gene expression in iPSCs differentiated as embryoid bodies for 21 days. Each experiment in b – d was repeated independently at least twice with similar results. Data and error bars in e and f indicate the means ± SD ( n = 3). Source data is provided as a Source Data file.

Techniques Used: Western Blot, Knock-In, Single-cell Analysis, Expressing, Gene Expression

other:

Article Title: Compartmentalization of the SUMO/RNF4 pathway by SLX4 drives DNA repair.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Deposited Data Raw data This paper Mendeley Dataset: https://doi.org/10.17632/c5h63ykhpn.1 Experimental Models: Cell Lines Flp-In T-REx 293 Invitrogen Cat# R78007; RRID:CVCL_U427 SLX4-null (RA3331/E6E7/hTERT) Gift from Dr. Agata Smogorzewska N/A SLX4-complemented (RA3331/E6E7/hTERT) Gift from Dr. Agata Smogorzewska N/A HEK293T Sigma-Aldrich Cat# 12022001; RRID:CVCL_0063 CL-HJL-0104-A KI HEK 293T Biocytogen N/A Oligonucleotides siGENOME Non-Targeting siRNA Control Pools Dharmacon_Horizon Discovery Cat# ID:D-001206-13-20 siGENOME Human SLX4 siRNA Dharmacon_Horizon Discovery Cat# ID:M-014895-01-0020 siGENOME Human RNF4 siRNA Dharmacon_Horizon Discovery Cat# ID:M-006557-03-0020 siGENOME Human UBE2I siRNA Dharmacon_Horizon Discovery Cat# ID:M-004910-00-0020 siGENOME Human SENP6 siRNA Dharmacon_Horizon Discovery Cat# ID:M-006044-01-0020 The oligonucleotides used in this study are listed in Table S2 N/A Recombinant DNA pCDNA5_FRT_TO_TurboID-mCherry-Cry2 Addgene Cat# 166504; http://n2t.net/addgene:166504; RRID:Addgene_166504 pOG44 Flp-Recombinase Expression Vector ThermoFisher Scientific Cat# V600520 pLenti-CMV-Blast-PIP-FUCCI Addgene Cat# 138715; http://n2t.net/addgene:138715; RRID:Addgene_138715 pCRISPaint-HaloTag-PuroR Addgene Cat# 80960; http://n2t.net/addgene:80960; RRID:Addgene_80960 Software and Algorithms OMERO OME Remote Objects software RRID:SCR_002629 Image Lab Software (Version 5.2.1) Bio-Rad RRID:SCR_014210 Biorender Software Science Suite Inc. RRID:SCR_018361 Prism 8 (Version 8.4.2) Graphpad Software RRID:SCR_002798 FIJI Open source RRID:SCR_002285 Xcalibur (Version 4.0) Thermo Scientific RRID:SCR_014593 MaxQuant (Version 1.6.10.43) Max-Planck-Institute of Biochemistry RRID:SCR_014485 Other CO2 Incubator C150 Binder Cat# 9040-0078 Sonicator, VibraCell- 72405 BioBlock scientific N/A KNF LABOPORT Mini Diaphragm Vacuum Pump N 811 in Pumps, Compressors Dominique Dutscher Cat# KNF_28002 Centrifuge Hettich Mikro 200 Grosseron N/A Mini-PROTEAN Tetra Vertical Electrophoresis Cell Bio-Rad Cat# 1658004 PowerPac HC High-Current Power Supply Bio-Rad Cat# 1645052 Trans-Blot Turbo Transfer System Bio-Rad Cat# 1704150 Slot Blot blotting manifold Hoefer Cat# PR648 Chemical Duty Pump, 220 V/50 Hz Merck Cat# WP6122050 Acclaim PepMap 100 C18 HPLC Columns ThermoFisher Scientific Cat# 164568 e4 Molecular Cell 83, 1640–1658.e1–e9, May 18, 2023

Article Title: The Degradation of Botulinum Neurotoxin Light Chains Using PROTACs.
Article Snippet: PROTACs were purchased from commercial sources: dTAGV-1, dTAG-13, and dTAGV1-NEG (Tocris Bioscience, Bristol, UK); HaloPROTAC3 (Promega, Madison, WI, USA).

Article Title: ZNF143 is a transcriptional regulator of nuclear-encoded mitochondrial genes that acts independently of looping and CTCF.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Anti-CTCF EMD Millipore Cat#07-729; RRID:AB_441965 Anti-TBP Abcam Cat#ab51841; RRID:AB_945758 Anti-ZNF143 Novus Cat#H00007702-M01, RRID:AB_2218328 Chemicals, peptides, and recombinant proteins 5-Ph-IAA MedChemExpress Cat#HY-134653 HaloPROTAC3 Promega Cat#GA3110 dTAG-13 Tocris Cat#6605 cOmplete protease inhibitor Roche Cat#11873580001 Micrococcal nuclease Worthington Biochemical Cat#LS004798 Critical commercial assays NEBNext Ultra II Library Prep Kit New England Biolabs Cat#E7645L Zymo ChIP DNA Clean and Concentrator Kit Zymo Research Cat#D5205 Zymo DNA Clean and Concentrator Kit Zymo Research Cat#D4034 Zymoclean Gel DNA Recovery Kit Zymo Research Cat#D4008 Deposited data fastSPT tracks, FRAP quantification, abundance quantification, DESeq2 outputs This paper Zenodo: https://doi.org/10.5281/zenodo.14056602 Micro-C, ChIP-seq, PRO-seq This paper GEO: GSE256246 Micro-C in mESCs Hsieh et al.74 GEO: GSE130275 ZNF143 ChIP-seq in HEK293T Ngondo-Mbongo et al.37 GEO: GSE39263 CTCF ChIP-seq in HEK293 ENCODE ENCODE: ENCSR617IFZ CTCF ChIP-seq in mESCs Hansen et al.14 GEO: GSE90994 CTCF ChIP-seq in mESCs ENCODE ENCODE: ENCSR000CCB CTCF ChIP-seq in mESCs ENCODE ENCODE: ENCSR362VNF PRO-seq in HEK293T Sathyan et al.41 GEO: GSE126919 H3K4me1 ChIP-seq in HEK293 ENCODE ENCODE: ENCSR000FCG H3K27ac ChIP-seq in HEK293 ENCODE ENCODE: ENCSR000FCH H3K4me1 ChIP-seq in mESCs ENCODE ENCODE: ENCSR032JUI H3K27ac ChIP-seq in mESCs Chronis et al.113 GEO: GSE90893 Experimental models: Cell lines Clone A, mouse embryonic stem cells, ZFP143-mAID2-HaloTag-V5, FLAG-SNAPfTagFKBP12(F36V)-CTCF, Rosa26-osTir1(F74G) This paper Available upon request Clone B, mouse embryonic stem cells, ZFP143-mAID2-HaloTag-V5, FLAG-SNAPfTagFKBP12(F36V)-CTCF, Rosa26-osTir1(F74G) This paper Available upon request Clone D, mouse embryonic stem cells, ZFP143SNAPfTag-FKBP12(F36V)-V5, FLAG-HaloTag-CTCF This paper Available upon request Clone 30, HEK293T cells, ZNF143-AID, GFP-ARF, osTir1 Sathyan et al.41 N/A HEK293T cells ATCC #CRL-3216; RRID:CVCL_0063 JM8.N4 mouse embryonic stem cells Pettitt et al.114 RRID:CVCL_J962 C87, mouse embryonic stem cells, FLAG-HaloTag-CTCF Hansen et al.14 Available upon request (Continued on next page) e1 Molecular Cell 85, 1–15.e1–e9, January 2, 2025

Article Title: The Degradation of Botulinum Neurotoxin Light Chains Using PROTACs
Article Snippet: PROTACs were purchased from commercial sources: dTAG V -1, dTAG-13, and dTAG V -1-NEG (Tocris Bioscience, Bristol, UK); HaloPROTAC3 (Promega, Madison, WI, USA).

Incubation:

Article Title: Putative Looping Factor ZNF143/ZFP143 is an Essential Transcriptional Regulator with No Looping Function
Article Snippet: The cells were passaged every 2–4 days by dissociation with TrypLE Express Enzyme (ThermoFisher #12605028). .. For depletion of ZFP143 in the clone A and clone B cell lines, the cells were incubated in media containing 100 μM 5-Ph-IAA (MedChemExpress #HY-134653) and 300 nM HaloPROTAC3 (Promega # GA3110), and for depletion of CTCF in the same lines the, the cells were incubated in media containing 100 nM dTAG-13 (Tocris # 6605). ..

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HaloPROTAC3 efficiently binds Halotag and HaloG3BP1 fusion proteins. ( A ) Chemical structure of HaloPROTAC3 and the enantiomeric form ent -HaloPROTAC3; the scheme indicates the warhead-binding VHL E3 ubiquitin ligase (VH285) and the chloroalkene moiety-binding Halotag (generated using Marvin JS 22.11.1, web-based chemical sketch tool by Chemaxon). ( B ) BV2 ΔG3BP1 + HaloG3 BP1 cells were treated with the indicated concentrations of HaloPROTAC3 for 48 h, and metabolic cell death was measured using CellTiter-Blue. Cell death was calculated as relative to the DMSO control, and CC 50 is indicated by a dotted line. ( C–E ) BV2 + Halotag, BV2 ΔG3BP1 + Halotag, or BV2 ΔG3BP1 + HaloG3 BP1 cells treated with the indicated concentrations of HaloPROTAC3 for 48 h were subjected to flow cytometry analysis after incubation for 2 h with the fluorescent ligand Halotag Janelia 647. ( C ) Gating strategy of Halotag + cells in the live cell population. ( D ) Representative histograms and ( E ) dose-response curves of the quantified percentage of Halotag + cells. The corresponding curve fitting parameters for each cell line are shown.

Journal: Antimicrobial Agents and Chemotherapy

Article Title: Proof of concept: targeted protein degradation of the stress granules component G3BP1 as an antiviral strategy against norovirus infection

doi: 10.1128/aac.01118-25

Figure Lengend Snippet: HaloPROTAC3 efficiently binds Halotag and HaloG3BP1 fusion proteins. ( A ) Chemical structure of HaloPROTAC3 and the enantiomeric form ent -HaloPROTAC3; the scheme indicates the warhead-binding VHL E3 ubiquitin ligase (VH285) and the chloroalkene moiety-binding Halotag (generated using Marvin JS 22.11.1, web-based chemical sketch tool by Chemaxon). ( B ) BV2 ΔG3BP1 + HaloG3 BP1 cells were treated with the indicated concentrations of HaloPROTAC3 for 48 h, and metabolic cell death was measured using CellTiter-Blue. Cell death was calculated as relative to the DMSO control, and CC 50 is indicated by a dotted line. ( C–E ) BV2 + Halotag, BV2 ΔG3BP1 + Halotag, or BV2 ΔG3BP1 + HaloG3 BP1 cells treated with the indicated concentrations of HaloPROTAC3 for 48 h were subjected to flow cytometry analysis after incubation for 2 h with the fluorescent ligand Halotag Janelia 647. ( C ) Gating strategy of Halotag + cells in the live cell population. ( D ) Representative histograms and ( E ) dose-response curves of the quantified percentage of Halotag + cells. The corresponding curve fitting parameters for each cell line are shown.

Article Snippet: HaloPROTAC3 (VH285-PEG4-C4-Cl, HY-111997) and VH032 (HY-120217) were purchased from MedChemExpress. ent -HaloPROTAC3 was obtained from Promega.

Techniques: Binding Assay, Ubiquitin Proteomics, Generated, Control, Flow Cytometry, Incubation

HaloPROTA3 degrades Halotag and HaloG3BP1 fusion proteins. ( A–E ) Protein samples from BV2 + Halotag, ΔG3BP1 + Halotag, or ΔG3BP1 + HaloG3 BP1 cells treated for either 24 h or 48 h with HaloPROTAC3 or ent -HaloPROTAC3 were subjected to Western blot analysis using antibodies against Halotag, G3BP1, or GAPDH. ( A ) Representative Western blots and ( B ) quantification of the Halotag protein expression in cells treated with HaloPROTAC3 normalized to GAPDH. ( C ) BV2 ΔG3BP1 + HaloG3 BP1 cells treated for 48 h with either 1.2 µM of HaloPROTAC3 or the indicated concentrations of ent -HaloPROTAC3 were subjected to flow cytometry analysis, as stated in . Data is expressed as mean ± standard error of the mean with ∗∗∗∗ indicating P ≤ 0.0001. ( D ) Representative Western blots and ( E ) quantification of the Halotag protein expression in cells treated with ent- HaloPROTAC3 normalized to GAPDH.

Journal: Antimicrobial Agents and Chemotherapy

Article Title: Proof of concept: targeted protein degradation of the stress granules component G3BP1 as an antiviral strategy against norovirus infection

doi: 10.1128/aac.01118-25

Figure Lengend Snippet: HaloPROTA3 degrades Halotag and HaloG3BP1 fusion proteins. ( A–E ) Protein samples from BV2 + Halotag, ΔG3BP1 + Halotag, or ΔG3BP1 + HaloG3 BP1 cells treated for either 24 h or 48 h with HaloPROTAC3 or ent -HaloPROTAC3 were subjected to Western blot analysis using antibodies against Halotag, G3BP1, or GAPDH. ( A ) Representative Western blots and ( B ) quantification of the Halotag protein expression in cells treated with HaloPROTAC3 normalized to GAPDH. ( C ) BV2 ΔG3BP1 + HaloG3 BP1 cells treated for 48 h with either 1.2 µM of HaloPROTAC3 or the indicated concentrations of ent -HaloPROTAC3 were subjected to flow cytometry analysis, as stated in . Data is expressed as mean ± standard error of the mean with ∗∗∗∗ indicating P ≤ 0.0001. ( D ) Representative Western blots and ( E ) quantification of the Halotag protein expression in cells treated with ent- HaloPROTAC3 normalized to GAPDH.

Article Snippet: HaloPROTAC3 (VH285-PEG4-C4-Cl, HY-111997) and VH032 (HY-120217) were purchased from MedChemExpress. ent -HaloPROTAC3 was obtained from Promega.

Techniques: Western Blot, Expressing, Flow Cytometry

Degradation of G3BP1 using HaloPROTAC3 shows antiviral activity against norovirus. BV2 ΔG3BP1 + HaloG3 BP1 cells were treated for 48 h with the shown concentrations of HaloPROTA3 and infected with MNV1. After 1 h of virus adsorption, the virus inoculum was removed, and HaloPROTAC3 was re-added to the cell culture medium for the remaining infection time. ( A ) Scheme of the experimental setup of the antiviral assays. ( B–D ) Cells infected with MOI 1 TCID 50 /cell for 16 h were incubated with fluorescent ligand Halotag Janelia 647 for 2 h, harvested, and stained with anti-dsRNA antibody to assess the percentage of infection. Cells were then subjected to flow cytometry analysis. ( B ) Gating strategy showing first the duplet exclusion for selection of single cells, followed by dead cell exclusion within the single-cell population. Halotag + cells and dsRNA + cells were gated exclusively from the live cell population. ( C ) Quantification of the percentage of Halotag + and ( D ) dsRNA + (infected) cells across HaloPROTAC3 concentrations. ( E ) Total RNA was isolated from cells infected with MOI 1 TCID 50 /cell for 10 h and used to quantify the levels of viral RNA (expressed as vRNA copies/µg RNA, normalized to the DMSO control), by RT-qPCR. ( F ) Extracellular infectious virus yield in supernatants from cells infected with MOI 1 TCID 50 /cell for 16 h and treated with the indicated concentrations of HaloPROTAC3 was quantified by TCID 50 in BV2 cells. ( G, H ) Infection-induced cell death was detected using the NIR dye measured in the live-cell imaging IncuCyte system. ( G ) Representative pictures after 24 h.p.i.; NIR + cells are shown in blue. ( H ) Quantification of the total number of NIR + cells for the shown HaloPROTAC3 concentrations. Data is expressed as mean ± standard error of the mean with ∗ , ∗∗ , ∗∗∗ , and ∗∗∗∗ indicating P ≤ 0.05, P ≤ 0.01, P ≤ 0.001, and P ≤ 0.0001 respectively.

Journal: Antimicrobial Agents and Chemotherapy

Article Title: Proof of concept: targeted protein degradation of the stress granules component G3BP1 as an antiviral strategy against norovirus infection

doi: 10.1128/aac.01118-25

Figure Lengend Snippet: Degradation of G3BP1 using HaloPROTAC3 shows antiviral activity against norovirus. BV2 ΔG3BP1 + HaloG3 BP1 cells were treated for 48 h with the shown concentrations of HaloPROTA3 and infected with MNV1. After 1 h of virus adsorption, the virus inoculum was removed, and HaloPROTAC3 was re-added to the cell culture medium for the remaining infection time. ( A ) Scheme of the experimental setup of the antiviral assays. ( B–D ) Cells infected with MOI 1 TCID 50 /cell for 16 h were incubated with fluorescent ligand Halotag Janelia 647 for 2 h, harvested, and stained with anti-dsRNA antibody to assess the percentage of infection. Cells were then subjected to flow cytometry analysis. ( B ) Gating strategy showing first the duplet exclusion for selection of single cells, followed by dead cell exclusion within the single-cell population. Halotag + cells and dsRNA + cells were gated exclusively from the live cell population. ( C ) Quantification of the percentage of Halotag + and ( D ) dsRNA + (infected) cells across HaloPROTAC3 concentrations. ( E ) Total RNA was isolated from cells infected with MOI 1 TCID 50 /cell for 10 h and used to quantify the levels of viral RNA (expressed as vRNA copies/µg RNA, normalized to the DMSO control), by RT-qPCR. ( F ) Extracellular infectious virus yield in supernatants from cells infected with MOI 1 TCID 50 /cell for 16 h and treated with the indicated concentrations of HaloPROTAC3 was quantified by TCID 50 in BV2 cells. ( G, H ) Infection-induced cell death was detected using the NIR dye measured in the live-cell imaging IncuCyte system. ( G ) Representative pictures after 24 h.p.i.; NIR + cells are shown in blue. ( H ) Quantification of the total number of NIR + cells for the shown HaloPROTAC3 concentrations. Data is expressed as mean ± standard error of the mean with ∗ , ∗∗ , ∗∗∗ , and ∗∗∗∗ indicating P ≤ 0.05, P ≤ 0.01, P ≤ 0.001, and P ≤ 0.0001 respectively.

Article Snippet: HaloPROTAC3 (VH285-PEG4-C4-Cl, HY-111997) and VH032 (HY-120217) were purchased from MedChemExpress. ent -HaloPROTAC3 was obtained from Promega.

Techniques: Activity Assay, Infection, Virus, Adsorption, Cell Culture, Incubation, Staining, Flow Cytometry, Selection, Single Cell, Isolation, Control, Quantitative RT-PCR, Live Cell Imaging

Degradation of G3BP1 is required for HaloPROTAC3 antiviral activity. ( A, B ) BV2 ΔG3BP1 + HaloG3 BP1 cells were treated for 48 h with 1.2 µM of HaloPROTAC3 or ent -HaloPROTAC3, as indicated, and infected with MNV1. After 1 h of virus adsorption, the virus inoculum was removed and PROTACs re-added to the cell culture medium. ( A ) Extracellular infectious virus yield in supernatants from cells infected with MOI 1 TCID 50 /cell for 16 h and treated with either HaloPROTAC3 or ent -HALOPROTAC3 was quantified by TCID 50 in BV2 cells. ( B ) Infection-induced cell death was detected using the NIR dye measured in the live-cell imaging IncuCyte system. Shown is the quantification of the total number of NIR + cells for cells treated with either HaloPROTAC3 or ent -HaloPROTACs, measured every 3 h and followed for a total of 72 h.p.i. ( C–F ) BV2 ΔG3BP1 + HaloG3 BP1 cells were concomitantly treated for 48 h with 1.2 µM of HaloPROTAC3 and VH032 in the indicated concentrations. ( C ) Metabolic cell death was measured using CellTiter-Blue. Cell death was calculated as relative to the DMSO control. A concentration of 75 µM of VH032 is indicated by a dotted line. ( D ) Protein samples from cells treated for either 24 or 48 h with the indicated concentrations of HaloPROTAC3 and VH032 were subjected to Western blot analysis using antibodies directed Halotag or GAPDH. Bar plot shows the quantification of the Halotag protein bands from three independent experiments for all the experimental conditions. ( E ) Cells were infected with MOI 1 TCID 50 /cell of MNV1. After 1 h of virus adsorption, the virus inoculum was removed and drugs re-added to the cell culture medium. Infection-induced cell death was detected using the NIR dye measured in the live-cell imaging IncuCyte system, as stated in B . Shown is the quantification of the total number of NIR + cells measured after 24 h.p.i. ( F ) Extracellular infectious virus yield quantified by TCID 50 in BV2 cells from supernatants of cells infected with MOI 1 TCID 50 /cell for 16 h and treated with 1.2 µM of HaloPROTAC3 and 30 µM of VH032. Data is expressed as mean ± standard error of the mean with ∗∗ , ∗∗∗ , and ∗∗∗∗ indicating P ≤ 0.01, P ≤ 0.001, and P ≤ 0.0001 respectively. ‘ns’ denotes non-significant.

Journal: Antimicrobial Agents and Chemotherapy

Article Title: Proof of concept: targeted protein degradation of the stress granules component G3BP1 as an antiviral strategy against norovirus infection

doi: 10.1128/aac.01118-25

Figure Lengend Snippet: Degradation of G3BP1 is required for HaloPROTAC3 antiviral activity. ( A, B ) BV2 ΔG3BP1 + HaloG3 BP1 cells were treated for 48 h with 1.2 µM of HaloPROTAC3 or ent -HaloPROTAC3, as indicated, and infected with MNV1. After 1 h of virus adsorption, the virus inoculum was removed and PROTACs re-added to the cell culture medium. ( A ) Extracellular infectious virus yield in supernatants from cells infected with MOI 1 TCID 50 /cell for 16 h and treated with either HaloPROTAC3 or ent -HALOPROTAC3 was quantified by TCID 50 in BV2 cells. ( B ) Infection-induced cell death was detected using the NIR dye measured in the live-cell imaging IncuCyte system. Shown is the quantification of the total number of NIR + cells for cells treated with either HaloPROTAC3 or ent -HaloPROTACs, measured every 3 h and followed for a total of 72 h.p.i. ( C–F ) BV2 ΔG3BP1 + HaloG3 BP1 cells were concomitantly treated for 48 h with 1.2 µM of HaloPROTAC3 and VH032 in the indicated concentrations. ( C ) Metabolic cell death was measured using CellTiter-Blue. Cell death was calculated as relative to the DMSO control. A concentration of 75 µM of VH032 is indicated by a dotted line. ( D ) Protein samples from cells treated for either 24 or 48 h with the indicated concentrations of HaloPROTAC3 and VH032 were subjected to Western blot analysis using antibodies directed Halotag or GAPDH. Bar plot shows the quantification of the Halotag protein bands from three independent experiments for all the experimental conditions. ( E ) Cells were infected with MOI 1 TCID 50 /cell of MNV1. After 1 h of virus adsorption, the virus inoculum was removed and drugs re-added to the cell culture medium. Infection-induced cell death was detected using the NIR dye measured in the live-cell imaging IncuCyte system, as stated in B . Shown is the quantification of the total number of NIR + cells measured after 24 h.p.i. ( F ) Extracellular infectious virus yield quantified by TCID 50 in BV2 cells from supernatants of cells infected with MOI 1 TCID 50 /cell for 16 h and treated with 1.2 µM of HaloPROTAC3 and 30 µM of VH032. Data is expressed as mean ± standard error of the mean with ∗∗ , ∗∗∗ , and ∗∗∗∗ indicating P ≤ 0.01, P ≤ 0.001, and P ≤ 0.0001 respectively. ‘ns’ denotes non-significant.

Article Snippet: HaloPROTAC3 (VH285-PEG4-C4-Cl, HY-111997) and VH032 (HY-120217) were purchased from MedChemExpress. ent -HaloPROTAC3 was obtained from Promega.

Techniques: Activity Assay, Infection, Virus, Adsorption, Cell Culture, Live Cell Imaging, Control, Concentration Assay, Western Blot

Journal: Nature Communications

Article Title: Systematic comparison and base-editing-mediated directed protein evolution and functional screening yield superior auxin-inducible degron technology

doi: 10.1038/s41467-025-61848-1

Figure Lengend Snippet: a Schematics illustrate five degron technologies for ligand-inducible protein depletion. b , c Western blots indicate comparative depletion of endogenously tagged CTCF and RAD21 proteins in KOLF2.2J iPSCs. The endogenous genes had a homozygous knock-in of mAID, miniIAA7, FKBP12(F36V), HaloTag, and IKZF3 degron tags at the C-termini of the target genes, and the target proteins were induced to degrade by treating cells with 1 μM 5-Phenyl-indole-3-acetic acid (5-Ph-IAA, synthetic auxin), 500 μM Indole-3-acetic acid (IAA, auxin), 1 μM dTAG13, 1 μM HaloPROTAC3, or 1 μM Pomalidomide, respectively, for 0, 1, 6, and 24 h. d Western blots show relative recovery rates of endogenous CTCF protein levels in five degron systems. KOLF2.2J iPSCs were treated with respective ligands for 6 h, then the ligands were washed out, and target protein levels were assessed after 24 h and 48 h with western blot. e Bar plots show quantification of target protein depletion after 1 h, 6 h, and 24 h of treatment with each chemical degrader. f The bar plot shows relative CTCF protein recovery after ligand washout in each degron system. g The single cell analysis (UMAP visualization) shows the relative expression levels of endogenous VHL , CRBN genes, and CAG-promoter driven OsTIR1 gene expression in iPSCs differentiated as embryoid bodies for 21 days. Each experiment in b – d was repeated independently at least twice with similar results. Data and error bars in e and f indicate the means ± SD ( n = 3). Source data is provided as a Source Data file.

Article Snippet: At 48 h after seeding, cells were treated with 500 μM IAA (Millipore Sigma, 87-51-4), 1 μM 5-Ph-IAA, 1 μM dTAG13 (MedChem Express, HY-114421), 1 μM HaloPROTAC3 (Promega, GA3110) and Pomalidomide (Selleck Chemicals, S1567), respectively.

Techniques: Western Blot, Knock-In, Single-cell Analysis, Expressing, Gene Expression

Journal: Nature Communications

Article Title: Systematic comparison and base-editing-mediated directed protein evolution and functional screening yield superior auxin-inducible degron technology

doi: 10.1038/s41467-025-61848-1

Techniques: Western Blot, Knock-In, Single-cell Analysis, Expressing, Gene Expression

Review

haloprotac3 (Promega)

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