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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/product/full+length+wt+polq/bio_rxiv__2025__11__26__690880-79-5-10?v=Addgene+inc
    Average 93 stars, based on 9 article reviews
    full length wt polq - by Bioz Stars, 2026-08
    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



    Similar Products

    wild type wt full length polq cdna  (New England Biolabs)
    86
    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/product/full+length+wt+polq/pmc08224818-297-0-21?v=New+England+Biolabs
    Average 86 stars, based on 1 article reviews
    wild type wt full length polq cdna - by Bioz Stars, 2026-08
    86/100 stars
    		  Buy from Supplier
    full length wt polq  (Addgene inc)
    93
    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
    https://www.bioz.com/product/full+length+wt+polq/bio_rxiv__2025__11__26__690880-79-5-10?v=Addgene+inc
    Average 93 stars, based on 1 article reviews
    full length wt polq - by Bioz Stars, 2026-08
    93/100 stars
    		  Buy from Supplier

    Image Search Results


    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

    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