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full length wt polq  (Addgene inc)


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    Structured Review

    Addgene inc full length wt polq
    Variants display higher affinity for CPD damaged DNA compared to WT Pol θ. WT <t>POLQ</t> and L2538R, E2406K, and T2161I variants were titrated from 0-1800 nM against 10 nM undamaged (circles) or damaged (triangles) dsDNA. Bound and unbound products were separated on a 6% non-denaturing gel and quantified using ImageQuant software. K D(DNA) was calculated using (Methods).
    Full Length Wt Polq, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/full length wt polq/product/Addgene inc
    Average 93 stars, based on 9 article reviews
    full length wt polq - by Bioz Stars, 2026-06
    93/100 stars

    Images

    1) Product Images from "POLQ variants with aberrant DNA polymerase activity protect against UV-induced cell death"

    Article Title: POLQ variants with aberrant DNA polymerase activity protect against UV-induced cell death

    Journal: bioRxiv

    doi: 10.1101/2025.11.26.690880

    Variants display higher affinity for CPD damaged DNA compared to WT Pol θ. WT POLQ and L2538R, E2406K, and T2161I variants were titrated from 0-1800 nM against 10 nM undamaged (circles) or damaged (triangles) dsDNA. Bound and unbound products were separated on a 6% non-denaturing gel and quantified using ImageQuant software. K D(DNA) was calculated using (Methods).
    Figure Legend Snippet: Variants display higher affinity for CPD damaged DNA compared to WT Pol θ. WT POLQ and L2538R, E2406K, and T2161I variants were titrated from 0-1800 nM against 10 nM undamaged (circles) or damaged (triangles) dsDNA. Bound and unbound products were separated on a 6% non-denaturing gel and quantified using ImageQuant software. K D(DNA) was calculated using (Methods).

    Techniques Used: Software


    Figure Legend Snippet:

    Techniques Used:

    WT Pol θ and cancer variants bypass and extend past CPD damaged DNA. A representative denaturing gel demonstrates primer extension of CPD dsDNA. Under single-turnover conditions 750 nM of WT POLQ and L2538R, E2406K, and T2161I variants were preincubated with 50 nM of CPD damaged dsDNA substrate and then combined with 10 mM MgCl 2 and either no dNTPs or 50 µM of All dNTPs, dATP, dCTP, dGTP, or dTTP for 5 minutes at 37°C. DNA extension products were separated on a denaturing gel and visualized on a Typhoon scanner. Each n+1 band represents extension past the CPD damage with either correct (dATP) or incorrect incorporation (dCTP, dGTP, or dTTP). Each additional band represents another extension with a maximum product of n+12.
    Figure Legend Snippet: WT Pol θ and cancer variants bypass and extend past CPD damaged DNA. A representative denaturing gel demonstrates primer extension of CPD dsDNA. Under single-turnover conditions 750 nM of WT POLQ and L2538R, E2406K, and T2161I variants were preincubated with 50 nM of CPD damaged dsDNA substrate and then combined with 10 mM MgCl 2 and either no dNTPs or 50 µM of All dNTPs, dATP, dCTP, dGTP, or dTTP for 5 minutes at 37°C. DNA extension products were separated on a denaturing gel and visualized on a Typhoon scanner. Each n+1 band represents extension past the CPD damage with either correct (dATP) or incorrect incorporation (dCTP, dGTP, or dTTP). Each additional band represents another extension with a maximum product of n+12.

    Techniques Used:


    Figure Legend Snippet:

    Techniques Used:

    WT POLQ and variants E2406K, and T2161I reduce the number of double strand brakes in UV treated cells. MCF7 cells melanocytes were co-transfected with either a scrambled control siRNA oligo or an siRNA oligo to POLQ and either a siRNA resistant WT, L2538R, E2406K, or T2161I POLQ rescue plasmid. Cells were allowed to grow for 48-72 hrs. and then treated with either 5J ultra-violet radiation or not and allowed to recover for 24 hrs. Neutral comet assays were performed using Comet slides (n ≥ 50 cells) and analyzed using CometScore software. Error bars represent standard deviation, and significance was determined using One-way ANOVA (**** p < 0.0001).
    Figure Legend Snippet: WT POLQ and variants E2406K, and T2161I reduce the number of double strand brakes in UV treated cells. MCF7 cells melanocytes were co-transfected with either a scrambled control siRNA oligo or an siRNA oligo to POLQ and either a siRNA resistant WT, L2538R, E2406K, or T2161I POLQ rescue plasmid. Cells were allowed to grow for 48-72 hrs. and then treated with either 5J ultra-violet radiation or not and allowed to recover for 24 hrs. Neutral comet assays were performed using Comet slides (n ≥ 50 cells) and analyzed using CometScore software. Error bars represent standard deviation, and significance was determined using One-way ANOVA (**** p < 0.0001).

    Techniques Used: Transfection, Control, Plasmid Preparation, Software, Standard Deviation



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    full length wt polq  (Addgene inc)
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    Addgene inc full length wt polq
    Variants display higher affinity for CPD damaged DNA compared to WT Pol θ. WT <t>POLQ</t> and L2538R, E2406K, and T2161I variants were titrated from 0-1800 nM against 10 nM undamaged (circles) or damaged (triangles) dsDNA. Bound and unbound products were separated on a 6% non-denaturing gel and quantified using ImageQuant software. K D(DNA) was calculated using (Methods).
    Full Length Wt Polq, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 93 stars, based on 1 article reviews
    full length wt polq - by Bioz Stars, 2026-06
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    wild type wt full length polq cdna  (New England Biolabs)
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    New England Biolabs wild type wt full length polq cdna
    a . A schematic shows how the PARPi resistant clones R1-R4 were generated. b . A Western blot of WT and BRCA1 −/− RPE1 cells and the PARPi resistant clones, using an anti-BRCA1 antibody (Millipore #OP92). No BRCA1 reversion was observed in the clones. c-d. NVB sensitivity of WT RPE1, BRCA1 −/− RPE1, and PARPi-resistant BRCA1 −/− clones (R1-R4). R1 was selected by olaparib and R2-R4 were selected by Niraparib. R1 and R2 were tested in clonogenic survival assays (12–14 days) (c) . R3 and R4 were tested in CellTiter-Glo cell viability assay (6 days) ( d ). Data are mean ± SEM, n = 3 independent cultures. e . <t>POLQ</t> expression at mRNA level and protein level in WT RPE1, BRCA1 −/− RPE1, and PARPi-resistant BRCA1 −/− clones (R1-R4). POLQ mRNA was measured by qRT-PCR and normalized to beta-Actin (ACTB). Mean ± SD of n=4 independent cultures. f . POLQ expression at mRNA level in parental (P), PARPi resistant (R), and empty vector (+EV) or <t>BRCA1-cDNA</t> (+BR1) complemented MDA-MB-436 cells. Mean ± SD of n=4 independent cultures. g . POLQ expression at mRNA level in PDX models DF53, DF83 and DF149. Mean of 4 technical replicates for each PDX model are shown. h . Olaparib sensitivity of CAPAN1 (BRCA2 mutated) and CAPAN1-CR cells (BRCA2 CRISPR edited back to wild type) in clonogenic survival assays. i . NVB sensitivity of CAPAN1 and CAPAN1-CR cells in clonogenic survival assays. Mean of n=2 independent experiments are shown in h and i . Statistical analysis in h and i were paired t-test, * p < 0.05, j . The expression levels of POLQ in CAPAN1 and CAPAN1-CR cells, analyzed by Western blotting.
    Wild Type Wt Full Length Polq Cdna, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/wild type wt full length polq cdna/product/New England Biolabs
    Average 86 stars, based on 1 article reviews
    wild type wt full length polq cdna - by Bioz Stars, 2026-06
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    Image Search Results


    Variants display higher affinity for CPD damaged DNA compared to WT Pol θ. WT POLQ and L2538R, E2406K, and T2161I variants were titrated from 0-1800 nM against 10 nM undamaged (circles) or damaged (triangles) dsDNA. Bound and unbound products were separated on a 6% non-denaturing gel and quantified using ImageQuant software. K D(DNA) was calculated using (Methods).

    Journal: bioRxiv

    Article Title: POLQ variants with aberrant DNA polymerase activity protect against UV-induced cell death

    doi: 10.1101/2025.11.26.690880

    Figure Lengend Snippet: Variants display higher affinity for CPD damaged DNA compared to WT Pol θ. WT POLQ and L2538R, E2406K, and T2161I variants were titrated from 0-1800 nM against 10 nM undamaged (circles) or damaged (triangles) dsDNA. Bound and unbound products were separated on a 6% non-denaturing gel and quantified using ImageQuant software. K D(DNA) was calculated using (Methods).

    Article Snippet: WT POLQ rescue plasmids contained full-length WT POLQ obtained from Addgene (Addgene plasmid # 73132; http://n2t.net/addgene:73132 ; RRID:Addgene_73132, [ ]).

    Techniques: Software

    Journal: bioRxiv

    Article Title: POLQ variants with aberrant DNA polymerase activity protect against UV-induced cell death

    doi: 10.1101/2025.11.26.690880

    Figure Lengend Snippet:

    Article Snippet: WT POLQ rescue plasmids contained full-length WT POLQ obtained from Addgene (Addgene plasmid # 73132; http://n2t.net/addgene:73132 ; RRID:Addgene_73132, [ ]).

    Techniques:

    WT Pol θ and cancer variants bypass and extend past CPD damaged DNA. A representative denaturing gel demonstrates primer extension of CPD dsDNA. Under single-turnover conditions 750 nM of WT POLQ and L2538R, E2406K, and T2161I variants were preincubated with 50 nM of CPD damaged dsDNA substrate and then combined with 10 mM MgCl 2 and either no dNTPs or 50 µM of All dNTPs, dATP, dCTP, dGTP, or dTTP for 5 minutes at 37°C. DNA extension products were separated on a denaturing gel and visualized on a Typhoon scanner. Each n+1 band represents extension past the CPD damage with either correct (dATP) or incorrect incorporation (dCTP, dGTP, or dTTP). Each additional band represents another extension with a maximum product of n+12.

    Journal: bioRxiv

    Article Title: POLQ variants with aberrant DNA polymerase activity protect against UV-induced cell death

    doi: 10.1101/2025.11.26.690880

    Figure Lengend Snippet: WT Pol θ and cancer variants bypass and extend past CPD damaged DNA. A representative denaturing gel demonstrates primer extension of CPD dsDNA. Under single-turnover conditions 750 nM of WT POLQ and L2538R, E2406K, and T2161I variants were preincubated with 50 nM of CPD damaged dsDNA substrate and then combined with 10 mM MgCl 2 and either no dNTPs or 50 µM of All dNTPs, dATP, dCTP, dGTP, or dTTP for 5 minutes at 37°C. DNA extension products were separated on a denaturing gel and visualized on a Typhoon scanner. Each n+1 band represents extension past the CPD damage with either correct (dATP) or incorrect incorporation (dCTP, dGTP, or dTTP). Each additional band represents another extension with a maximum product of n+12.

    Article Snippet: WT POLQ rescue plasmids contained full-length WT POLQ obtained from Addgene (Addgene plasmid # 73132; http://n2t.net/addgene:73132 ; RRID:Addgene_73132, [ ]).

    Techniques:

    WT POLQ and variants E2406K, and T2161I reduce the number of double strand brakes in UV treated cells. MCF7 cells melanocytes were co-transfected with either a scrambled control siRNA oligo or an siRNA oligo to POLQ and either a siRNA resistant WT, L2538R, E2406K, or T2161I POLQ rescue plasmid. Cells were allowed to grow for 48-72 hrs. and then treated with either 5J ultra-violet radiation or not and allowed to recover for 24 hrs. Neutral comet assays were performed using Comet slides (n ≥ 50 cells) and analyzed using CometScore software. Error bars represent standard deviation, and significance was determined using One-way ANOVA (**** p < 0.0001).

    Journal: bioRxiv

    Article Title: POLQ variants with aberrant DNA polymerase activity protect against UV-induced cell death

    doi: 10.1101/2025.11.26.690880

    Figure Lengend Snippet: WT POLQ and variants E2406K, and T2161I reduce the number of double strand brakes in UV treated cells. MCF7 cells melanocytes were co-transfected with either a scrambled control siRNA oligo or an siRNA oligo to POLQ and either a siRNA resistant WT, L2538R, E2406K, or T2161I POLQ rescue plasmid. Cells were allowed to grow for 48-72 hrs. and then treated with either 5J ultra-violet radiation or not and allowed to recover for 24 hrs. Neutral comet assays were performed using Comet slides (n ≥ 50 cells) and analyzed using CometScore software. Error bars represent standard deviation, and significance was determined using One-way ANOVA (**** p < 0.0001).

    Article Snippet: WT POLQ rescue plasmids contained full-length WT POLQ obtained from Addgene (Addgene plasmid # 73132; http://n2t.net/addgene:73132 ; RRID:Addgene_73132, [ ]).

    Techniques: Transfection, Control, Plasmid Preparation, Software, Standard Deviation

    a . A schematic shows how the PARPi resistant clones R1-R4 were generated. b . A Western blot of WT and BRCA1 −/− RPE1 cells and the PARPi resistant clones, using an anti-BRCA1 antibody (Millipore #OP92). No BRCA1 reversion was observed in the clones. c-d. NVB sensitivity of WT RPE1, BRCA1 −/− RPE1, and PARPi-resistant BRCA1 −/− clones (R1-R4). R1 was selected by olaparib and R2-R4 were selected by Niraparib. R1 and R2 were tested in clonogenic survival assays (12–14 days) (c) . R3 and R4 were tested in CellTiter-Glo cell viability assay (6 days) ( d ). Data are mean ± SEM, n = 3 independent cultures. e . POLQ expression at mRNA level and protein level in WT RPE1, BRCA1 −/− RPE1, and PARPi-resistant BRCA1 −/− clones (R1-R4). POLQ mRNA was measured by qRT-PCR and normalized to beta-Actin (ACTB). Mean ± SD of n=4 independent cultures. f . POLQ expression at mRNA level in parental (P), PARPi resistant (R), and empty vector (+EV) or BRCA1-cDNA (+BR1) complemented MDA-MB-436 cells. Mean ± SD of n=4 independent cultures. g . POLQ expression at mRNA level in PDX models DF53, DF83 and DF149. Mean of 4 technical replicates for each PDX model are shown. h . Olaparib sensitivity of CAPAN1 (BRCA2 mutated) and CAPAN1-CR cells (BRCA2 CRISPR edited back to wild type) in clonogenic survival assays. i . NVB sensitivity of CAPAN1 and CAPAN1-CR cells in clonogenic survival assays. Mean of n=2 independent experiments are shown in h and i . Statistical analysis in h and i were paired t-test, * p < 0.05, j . The expression levels of POLQ in CAPAN1 and CAPAN1-CR cells, analyzed by Western blotting.

    Journal: Nature cancer

    Article Title: A first-in-class Polymerase Theta Inhibitor selectively targets Homologous-Recombination-Deficient Tumors

    doi: 10.1038/s43018-021-00203-x

    Figure Lengend Snippet: a . A schematic shows how the PARPi resistant clones R1-R4 were generated. b . A Western blot of WT and BRCA1 −/− RPE1 cells and the PARPi resistant clones, using an anti-BRCA1 antibody (Millipore #OP92). No BRCA1 reversion was observed in the clones. c-d. NVB sensitivity of WT RPE1, BRCA1 −/− RPE1, and PARPi-resistant BRCA1 −/− clones (R1-R4). R1 was selected by olaparib and R2-R4 were selected by Niraparib. R1 and R2 were tested in clonogenic survival assays (12–14 days) (c) . R3 and R4 were tested in CellTiter-Glo cell viability assay (6 days) ( d ). Data are mean ± SEM, n = 3 independent cultures. e . POLQ expression at mRNA level and protein level in WT RPE1, BRCA1 −/− RPE1, and PARPi-resistant BRCA1 −/− clones (R1-R4). POLQ mRNA was measured by qRT-PCR and normalized to beta-Actin (ACTB). Mean ± SD of n=4 independent cultures. f . POLQ expression at mRNA level in parental (P), PARPi resistant (R), and empty vector (+EV) or BRCA1-cDNA (+BR1) complemented MDA-MB-436 cells. Mean ± SD of n=4 independent cultures. g . POLQ expression at mRNA level in PDX models DF53, DF83 and DF149. Mean of 4 technical replicates for each PDX model are shown. h . Olaparib sensitivity of CAPAN1 (BRCA2 mutated) and CAPAN1-CR cells (BRCA2 CRISPR edited back to wild type) in clonogenic survival assays. i . NVB sensitivity of CAPAN1 and CAPAN1-CR cells in clonogenic survival assays. Mean of n=2 independent experiments are shown in h and i . Statistical analysis in h and i were paired t-test, * p < 0.05, j . The expression levels of POLQ in CAPAN1 and CAPAN1-CR cells, analyzed by Western blotting.

    Article Snippet: Wild type (WT) full length POLQ cDNA was assembled by PCR with a FLAG-P2A-BLAST cassette and subsequently cloned by Gibson assembly (New England BioLabs, #E2611L) in the PiggyBac PBCAG-eGFP vector (Addgene #40973) using BsrGI and SbfI sites to obtain the PiggyBac-eGFP-POLQ-FLAG-P2A-BLAST vector (referred here as GFP-Polθ WT).

    Techniques: Clone Assay, Generated, Western Blot, Viability Assay, Expressing, Quantitative RT-PCR, Plasmid Preparation, CRISPR

    a-b , Olaparib ( a ) and NVB ( b ) sensitivity of UWB1.289 and a PARPi resistant UWB1.289 clone (UWB1.289-YSR12) in CellTiter-Glo assays. Data shown are mean ± SD, n = 4 biological replicates. c . Western blot analysis of POLQ expression levels in in WT and BRCA1 −/− RPE1 cells and the PARPi resistant BRCA1 −/− clones. d. Olaparib sensitivity of PEO1 (BRCA2 mutated) and PEO4 cells (BRCA2 restored). e . NVB sensitivity of PEO1 and PEO4 cells. Data shown in d and e are Mean ± SD, n= 3 biological replicates. f . A Western blot shows POLQ expression in BRCA2-decient cells lines (PEO1) and their counterparts with BRCA2 reverted to wild type (PEO4).

    Journal: Nature cancer

    Article Title: A first-in-class Polymerase Theta Inhibitor selectively targets Homologous-Recombination-Deficient Tumors

    doi: 10.1038/s43018-021-00203-x

    Figure Lengend Snippet: a-b , Olaparib ( a ) and NVB ( b ) sensitivity of UWB1.289 and a PARPi resistant UWB1.289 clone (UWB1.289-YSR12) in CellTiter-Glo assays. Data shown are mean ± SD, n = 4 biological replicates. c . Western blot analysis of POLQ expression levels in in WT and BRCA1 −/− RPE1 cells and the PARPi resistant BRCA1 −/− clones. d. Olaparib sensitivity of PEO1 (BRCA2 mutated) and PEO4 cells (BRCA2 restored). e . NVB sensitivity of PEO1 and PEO4 cells. Data shown in d and e are Mean ± SD, n= 3 biological replicates. f . A Western blot shows POLQ expression in BRCA2-decient cells lines (PEO1) and their counterparts with BRCA2 reverted to wild type (PEO4).

    Article Snippet: Wild type (WT) full length POLQ cDNA was assembled by PCR with a FLAG-P2A-BLAST cassette and subsequently cloned by Gibson assembly (New England BioLabs, #E2611L) in the PiggyBac PBCAG-eGFP vector (Addgene #40973) using BsrGI and SbfI sites to obtain the PiggyBac-eGFP-POLQ-FLAG-P2A-BLAST vector (referred here as GFP-Polθ WT).

    Techniques: Western Blot, Expressing, Clone Assay