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t7ei digestion (New England Biolabs)

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New England Biolabs t7ei digestion
T7ei Digestion, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 2704 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/t7ei/T7+Endonuclease+I/pmc12678317-121-10-12
Average 99 stars, based on 2704 article reviews

t7ei digestion - by Bioz Stars, 2026-10

99/100 stars

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Expressing:

Article Title: Protocol to identify SINE-VNTR-Alu regulators using genome-wide screening in human K562 cells
Article Snippet: .. Cas9 is used for genome editing with the expression of sgRNA, and BFP is used to indicate Cas9 protein expression level. a. K562-Cas9-BFP cells should be cultured in RPMI 1640 medium containing 10% fetal bovine serum (FBS), 2 mM L-glutamine, 0.1 mM Non-Essential Amino Acids, and 1% penicillin-streptomycin, and cultured at 37°C with 5% CO 2 . b. Validate the Cas9 cleavage efficiency through the T7 Endonuclease I (T7E1) assay (NEB, Cat# M0302S) as follows: i. Infect the sgRNA lentivirus targeting a specific gene. ii. ..

Cell Culture:

other:

Article Title: Protocol for rolling circle amplification-based nanopore sequencing of eccDNA from cell and plasma samples.
Article Snippet: DNA sample+ NFW 25 μL NEBuffer 2 3 μL T7 Endonuclease I 2 μL Total 30 μL STAR Protocols 7, 104865, December 18, 2026 13 39.

Polymerase Chain Reaction:

Article Title: High genotoxicity of CRISPR/Cas9 versus limited efficacy of CRISPRi in chicken primordial germ cells
Article Snippet: The amplicons were subjected to Sanger sequencing, and sequencing traces were analyzed using the Inference of CRISPR Edits (ICE) software tool (v3.0, Synthego). .. Additionally, PCR products were digested with T7 Endonuclease I (T7EI; NEB M0302S) and analyzed by agarose gel electrophoresis to visualize indels. .. Total RNA was extracted from cultured cells, and cDNA was synthesized from 1 μg RNA using HiScript III RT SuperMix (Vazyme R323-01) according to the manufacturer’s protocol. qPCR was performed in triplicate using Taq Pro Universal SYBR qPCR Master Mix (Vazyme Q712-02) on a Bio-Rad CFX Connect Real-Time PCR Detection System.

Agarose Gel Electrophoresis:

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a Size distribution of iRGD-DC@CaP as determined by dynamic light scattering, and morphology of iRGD-DC@CaP as revealed by TEM images. Scale bar: 200 nm. b , c DOX and Cas9 release from iRGD-DC@CaP under different situations ( n = 3 biologically independent samples). Representative flow cytometry analysis images ( d ) and quantitative analysis ( e ) of cellular uptake in B16F10 cells following a 4 h incubation with iRGD-DC@CaP or controls ( n = 3 biologically independent samples). f Confocal laser scanning microscopy images of B16F10 cells following a 4 h incubation with iRGD-DC@CaP or controls. Cas9 was labeled with Alexa Fluor 647 (AF647; red), and the nuclei were stained with DAPI (blue). Scale bars: 50 µm. T7EI assay ( g ) and quantitative analysis ( h ) of indel frequencies of the Nt5e gene in B16F10 cells transfected with iRGD-DC CD73 @CaP or controls for 48 h ( n = 3 biologically independent samples). i Mutation rates of Nt5e examined by NGS analysis in B16F10 cells treated with iRGD-DC CD73 @CaP. j , k Western blot ( j ) and flow cytometry quantitative analysis ( k ) of CD73 protein expression in B16F10 cells after 72 h incubation with iRGD-DC CD73 @CaP or controls ( n = 3 biologically independent samples). l The ADO contents in the cell medium of B16F10 cells after 72 h incubation with iRGD-DC CD73 @CaP or controls detected by ELISA kits ( n = 3 biologically independent samples). The experiments for f and i were independently repeated three times with similar results. Data is presented as the mean ± SD. Statistical difference was assessed in ( e , h , j , k and l ) using one-way ANOVA with Dunnett’s multiple comparison test by comparing the iRGD-DC CD73 @CaP group with each of the other treatment groups. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. G1: saline (negative control); G2: Free Cas9 RNP; G3: iRGD-modified nanoparticles loaded with scrambled sgRNA (iRGD-DC Scr @CaP, negative control); G4: Lipofectamine™ CRISPRMAX™-mediated delivery of Cas9/sgCD73 (MAX Cas9/sgCD73, positive control); G5: nanoparticle without iRGD loaded with Cas9/sgCD73 (DC CD73 @CaP); G6: iRGD-modified nanoparticle loaded with Cas9/sgCD73 (iRGD-DC CD73 @CaP). The exact P value and Source data are provided as a Source Data file.

Journal: Nature Communications

Article Title: CD73 disruption with tumor-specific CRISPR/Cas9 nanoparticles reduces adenosine-mediated immunosuppression and amplifies response to chemotherapy

doi: 10.1038/s41467-026-77547-4

Figure Lengend Snippet: a Size distribution of iRGD-DC@CaP as determined by dynamic light scattering, and morphology of iRGD-DC@CaP as revealed by TEM images. Scale bar: 200 nm. b , c DOX and Cas9 release from iRGD-DC@CaP under different situations ( n = 3 biologically independent samples). Representative flow cytometry analysis images ( d ) and quantitative analysis ( e ) of cellular uptake in B16F10 cells following a 4 h incubation with iRGD-DC@CaP or controls ( n = 3 biologically independent samples). f Confocal laser scanning microscopy images of B16F10 cells following a 4 h incubation with iRGD-DC@CaP or controls. Cas9 was labeled with Alexa Fluor 647 (AF647; red), and the nuclei were stained with DAPI (blue). Scale bars: 50 µm. T7EI assay ( g ) and quantitative analysis ( h ) of indel frequencies of the Nt5e gene in B16F10 cells transfected with iRGD-DC CD73 @CaP or controls for 48 h ( n = 3 biologically independent samples). i Mutation rates of Nt5e examined by NGS analysis in B16F10 cells treated with iRGD-DC CD73 @CaP. j , k Western blot ( j ) and flow cytometry quantitative analysis ( k ) of CD73 protein expression in B16F10 cells after 72 h incubation with iRGD-DC CD73 @CaP or controls ( n = 3 biologically independent samples). l The ADO contents in the cell medium of B16F10 cells after 72 h incubation with iRGD-DC CD73 @CaP or controls detected by ELISA kits ( n = 3 biologically independent samples). The experiments for f and i were independently repeated three times with similar results. Data is presented as the mean ± SD. Statistical difference was assessed in ( e , h , j , k and l ) using one-way ANOVA with Dunnett’s multiple comparison test by comparing the iRGD-DC CD73 @CaP group with each of the other treatment groups. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. G1: saline (negative control); G2: Free Cas9 RNP; G3: iRGD-modified nanoparticles loaded with scrambled sgRNA (iRGD-DC Scr @CaP, negative control); G4: Lipofectamine™ CRISPRMAX™-mediated delivery of Cas9/sgCD73 (MAX Cas9/sgCD73, positive control); G5: nanoparticle without iRGD loaded with Cas9/sgCD73 (DC CD73 @CaP); G6: iRGD-modified nanoparticle loaded with Cas9/sgCD73 (iRGD-DC CD73 @CaP). The exact P value and Source data are provided as a Source Data file.

Article Snippet: T7 endonuclease I (T7EI) enzyme, EnGen Cas9 NLS S. pyogenes, HiScribe T7 High Yield RNA Synthesis Kit were purchased from New England BioLabs (NEB).

Techniques: Dynamic Light Scattering Assay, Morphology, Flow Cytometry, Analysis, Incubation, Confocal, Laser-Scanning Microscopy, Labeling, Staining, T7EI Assay, Transfection, Mutagenesis, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Comparison, Saline, Negative Control, Modification, Positive Control

a Fluorescence imaging of B16F10 tumor-bearing C57BL/6 mice following injection of iRGD-DC@CaP and controls ( n = 3 mice). b Quantitative analysis of AF647-Cas9 fluorescence from major organs in mice bearing B16F10 at 1 h after intravenous injection of iRGD-DC@CaP or controls ( n = 3 mice). “% ID/g” represents the percentage of the injected dose per gram of tissue. The exact P value: DC@CaP vs. iRGD-DC@CaP ** P = 0.0087. c Schematic illustration of B16F10 tumor therapy in vivo. Created in BioRender. Sun, X. (2026) https://BioRender.com/f4h4ymp . d Individual and average tumor growth curves of B16F10 following different treatments ( n = 5 mice). e Tumor weight and the tumor inhibition ratio following different treatments ( n = 5 mice). f Survival analysis of the mice following different treatments ( n = 6 mice). P values were determined by the log-rank test. g Indel frequency of Nt5e loci measured by T7EI assay in B16F10 tumor tissues with the indicated treatments. The experiment was independently repeated three times with similar results. h Western blotting and relative quantitative analysis of CD73 proteins in B16F10 tumor tissues with the indicated treatments ( n = 3 biologically independent samples). i The flow cytometry quantitative analysis of the CD73 expression on tumor cells (CD45 − ), endothelial cells (CD45 − CD31 + ), cancer-associated fibroblasts (CAFs) (CD45 − CD31 − GP38 + ), macrophages (CD45 + CD11b + F4/80 + ), DCs (CD45 + CD11c + MHC-II + ), Tregs (CD45 + CD4 + CD25 + Foxp3 + ) and MDSCs (CD45 + CD11b + Gr-1 + ) in B16F10 tumor tissues after indicated treatments ( n = 5 mice). j The ADO contents in the tumor after different treatments ( n = 5 mice). Data is presented as the mean ± SD. Statistical differences were assessed using one-way ANOVA in ( b , e , h , i and j ) or two-way ANOVA in ( d ) with Dunnett’s multiple comparison test by comparing the iRGD-DC CD73 @CaP group with each of the other treatment groups. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. G1: saline (negative control); G2: iRGD-modified nanoparticles loaded with scrambled sgRNA (iRGD-DC Scr @CaP, negative control); G3: iRGD-modified nanoparticle loaded with Cas9/sgCD73 (iRGD-DC CD73 @CaP). The exact P value and Source data are provided as a Source Data file.

Journal: Nature Communications

Article Title: CD73 disruption with tumor-specific CRISPR/Cas9 nanoparticles reduces adenosine-mediated immunosuppression and amplifies response to chemotherapy

doi: 10.1038/s41467-026-77547-4

Figure Lengend Snippet: a Fluorescence imaging of B16F10 tumor-bearing C57BL/6 mice following injection of iRGD-DC@CaP and controls ( n = 3 mice). b Quantitative analysis of AF647-Cas9 fluorescence from major organs in mice bearing B16F10 at 1 h after intravenous injection of iRGD-DC@CaP or controls ( n = 3 mice). “% ID/g” represents the percentage of the injected dose per gram of tissue. The exact P value: DC@CaP vs. iRGD-DC@CaP ** P = 0.0087. c Schematic illustration of B16F10 tumor therapy in vivo. Created in BioRender. Sun, X. (2026) https://BioRender.com/f4h4ymp . d Individual and average tumor growth curves of B16F10 following different treatments ( n = 5 mice). e Tumor weight and the tumor inhibition ratio following different treatments ( n = 5 mice). f Survival analysis of the mice following different treatments ( n = 6 mice). P values were determined by the log-rank test. g Indel frequency of Nt5e loci measured by T7EI assay in B16F10 tumor tissues with the indicated treatments. The experiment was independently repeated three times with similar results. h Western blotting and relative quantitative analysis of CD73 proteins in B16F10 tumor tissues with the indicated treatments ( n = 3 biologically independent samples). i The flow cytometry quantitative analysis of the CD73 expression on tumor cells (CD45 − ), endothelial cells (CD45 − CD31 + ), cancer-associated fibroblasts (CAFs) (CD45 − CD31 − GP38 + ), macrophages (CD45 + CD11b + F4/80 + ), DCs (CD45 + CD11c + MHC-II + ), Tregs (CD45 + CD4 + CD25 + Foxp3 + ) and MDSCs (CD45 + CD11b + Gr-1 + ) in B16F10 tumor tissues after indicated treatments ( n = 5 mice). j The ADO contents in the tumor after different treatments ( n = 5 mice). Data is presented as the mean ± SD. Statistical differences were assessed using one-way ANOVA in ( b , e , h , i and j ) or two-way ANOVA in ( d ) with Dunnett’s multiple comparison test by comparing the iRGD-DC CD73 @CaP group with each of the other treatment groups. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. G1: saline (negative control); G2: iRGD-modified nanoparticles loaded with scrambled sgRNA (iRGD-DC Scr @CaP, negative control); G3: iRGD-modified nanoparticle loaded with Cas9/sgCD73 (iRGD-DC CD73 @CaP). The exact P value and Source data are provided as a Source Data file.

Article Snippet: T7 endonuclease I (T7EI) enzyme, EnGen Cas9 NLS S. pyogenes, HiScribe T7 High Yield RNA Synthesis Kit were purchased from New England BioLabs (NEB).

Techniques: Fluorescence, Imaging, Injection, Analysis, Mouse Assay, Tissue, In Vivo, Inhibition, T7EI Assay, Western Blot, Flow Cytometry, Expressing, Comparison, Saline, Negative Control, Modification

a Schematic illustration of experimental design for 4T1 tumor therapy in vivo. b , c Average ( b ) and individual ( c ) tumor growth curves of 4T1 following different treatments ( n = 5 mice). d Tumor weight and tumor inhibition ratio following different treatments ( n = 5 mice). e Survival analysis of the mice following different treatments ( n = 6 mice). f Indel frequency of Nt5e loci measured by T7EI assay in 4T1 tumor tissues with the indicated treatments. The experiment was independently repeated three times with similar results. g Western blot and relative quantitative analysis of CD73 protein expression in 4T1 tumor tissues with the indicated treatments ( n = 3 biologically independent samples). h The ADO contents in the tumor after different treatments ( n = 5 mice). i–t Flow cytometry quantitative analysis of CD80 + CD86 + DCs ( i ), absolute number of CD8 + T cells ( j ), CD3 + CD8 + T cells ( k ), CD4 + CD25 + Foxp3 + Tregs ( l ), CD8 + T/Tregs ratio ( m ), CD3 − CD49b + NK cells ( n ), F4/80 + CD86 + cells ( o ), F4/80 + iNOS + cells ( p ), F4/80 + CD206 + cells ( q ), F4/80 + Arg1 + cells ( r ), F4/80 + CD86 + M1/F4/80 + CD206 + M2 ratio ( s ), and CD11b + Gr-1 + MDSCs ( t ) in tumor after different treatments ( n = 5 mice). u–w Contents of the IL-2 ( u ), TNF-α ( v ), and IFN-γ ( w ) in tumor tissues after different treatments ( n = 5 mice). Data is presented as the mean ± SD. Statistical differences were analyzed by two-way ANOVA with Dunnett’s multiple comparison test in ( b ), one-way ANOVA with Dunnett’s multiple comparison test in ( d , g , and h–w ) by comparing the iRGD-DC CD73 @CaP group with each of the other treatment groups. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. G1: saline (negative control); G2: iRGD-modified nanoparticles loaded with scrambled sgRNA (iRGD-DC Scr @CaP, negative control); G3: iRGD-modified nanoparticle loaded with Cas9/sgCD73 (iRGD-DC CD73 @CaP). The exact P value and Source data are provided as a Source Data file.

Journal: Nature Communications

Article Title: CD73 disruption with tumor-specific CRISPR/Cas9 nanoparticles reduces adenosine-mediated immunosuppression and amplifies response to chemotherapy

doi: 10.1038/s41467-026-77547-4

Figure Lengend Snippet: a Schematic illustration of experimental design for 4T1 tumor therapy in vivo. b , c Average ( b ) and individual ( c ) tumor growth curves of 4T1 following different treatments ( n = 5 mice). d Tumor weight and tumor inhibition ratio following different treatments ( n = 5 mice). e Survival analysis of the mice following different treatments ( n = 6 mice). f Indel frequency of Nt5e loci measured by T7EI assay in 4T1 tumor tissues with the indicated treatments. The experiment was independently repeated three times with similar results. g Western blot and relative quantitative analysis of CD73 protein expression in 4T1 tumor tissues with the indicated treatments ( n = 3 biologically independent samples). h The ADO contents in the tumor after different treatments ( n = 5 mice). i–t Flow cytometry quantitative analysis of CD80 + CD86 + DCs ( i ), absolute number of CD8 + T cells ( j ), CD3 + CD8 + T cells ( k ), CD4 + CD25 + Foxp3 + Tregs ( l ), CD8 + T/Tregs ratio ( m ), CD3 − CD49b + NK cells ( n ), F4/80 + CD86 + cells ( o ), F4/80 + iNOS + cells ( p ), F4/80 + CD206 + cells ( q ), F4/80 + Arg1 + cells ( r ), F4/80 + CD86 + M1/F4/80 + CD206 + M2 ratio ( s ), and CD11b + Gr-1 + MDSCs ( t ) in tumor after different treatments ( n = 5 mice). u–w Contents of the IL-2 ( u ), TNF-α ( v ), and IFN-γ ( w ) in tumor tissues after different treatments ( n = 5 mice). Data is presented as the mean ± SD. Statistical differences were analyzed by two-way ANOVA with Dunnett’s multiple comparison test in ( b ), one-way ANOVA with Dunnett’s multiple comparison test in ( d , g , and h–w ) by comparing the iRGD-DC CD73 @CaP group with each of the other treatment groups. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. G1: saline (negative control); G2: iRGD-modified nanoparticles loaded with scrambled sgRNA (iRGD-DC Scr @CaP, negative control); G3: iRGD-modified nanoparticle loaded with Cas9/sgCD73 (iRGD-DC CD73 @CaP). The exact P value and Source data are provided as a Source Data file.

Article Snippet: T7 endonuclease I (T7EI) enzyme, EnGen Cas9 NLS S. pyogenes, HiScribe T7 High Yield RNA Synthesis Kit were purchased from New England BioLabs (NEB).

Techniques: In Vivo, Inhibition, Analysis, T7EI Assay, Western Blot, Expressing, Flow Cytometry, Comparison, Saline, Negative Control, Modification

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t7ei digestion (New England Biolabs)

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