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pmbplbcas12a  (Addgene inc)


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    Structured Review

    Addgene inc pmbplbcas12a
    Pmbplbcas12a, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 32 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/lbcas12a/pMBP-LbCas12a+(Plasmid+%23113431)/pm41832076-328-25-26
    Average 93 stars, based on 32 article reviews
    pmbplbcas12a - by Bioz Stars, 2026-09
    93/100 stars

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    Related Articles

    Expressing:

    Article Title: Harnessing noncanonical crRNAs to improve functionality of Cas12a orthologs
    Article Snippet: The product plasmids were then transformed into Rosetta(DE3)pLysS Competent Cells (Millipore Sigma, Catalog #70956) following the manufacturer’s protocols. .. For LbCas12a, AsCas12a, FnCas12a, and MbCas12a, protein expression plasmids were obtained from Addgene and used directly for protein expression., , ..

    Article Title: Harnessing noncanonical crRNAs to improve functionality of Cas12a orthologs.
    Article Snippet: The product plasmids were then transformed into Rosetta(DE3)pLysS Competent Cells (Millipore Sigma, Catalog #70956) following the manufacturer’s protocols. .. For LbCas12a, AsCas12a, FnCas12a, and MbCas12a, protein expression plasmids were obtained from Addgene and used directly for protein expression.3,42,43 ..

    Article Title: Highly parallel profiling of the activities and specificities of Cas12a variants in human cells
    Article Snippet: .. AsCas12a (Addgene, #69982), LbCas12a (Addgene, #69988), FnCas12a (Addgene, #69976), Lb2Cas12a (Addgene, #69983) human expression plasmids were purchased from the non-profit plasmid repository Addgene. .. To prepare the lentiviral vectors, we removed the Cas9-encoding sequence from the lentiCas9-Blast plasmid (Addgene, #52962), and cloned Cas12a-encoding sequences between XbaI and BamHI.

    Article Title: Post-cleavage target residence determines asymmetry in non-homologous end joining of Cas12a-induced DNA double strand breaks.
    Article Snippet: .. The expression plasmids for SpCas9 (Cat# 42,230) and LbCas12a (Cat# 69,988) were originally obtained from Addgene. ..

    Purification:

    Article Title: Combining Multiplexed CRISPR/Cas9-Nickase and PARP Inhibitors Efficiently and Precisely Targets Cancer Cells
    Article Snippet: .. Purification of recombinant Cas9 WT , nCas9 D10A , and LbCas12a-ultra The pET-LbCas12a-ultra vector was constructed using the DNA sequence encoding LbCas12a-ultra (18), which was cloned into the pET28a+ vector (Addgene, #69864-3). pETCas9-HN was derived from p3s-Cas9-HN (Addgene, #104171, RRID: Addgene_104171). .. These vectors were transformed into the C3013 strain (NEB, #C3013I), and recombinant Cas9 WT or LbCas12a-ultra protein was expressed at 25°C for 4 h by addition of 0.5 mM IPTG, followed by purification using Ni-NTA agarose beads (Qiagen, #30210).

    Recombinant:

    Article Title: Combining Multiplexed CRISPR/Cas9-Nickase and PARP Inhibitors Efficiently and Precisely Targets Cancer Cells
    Article Snippet: .. Purification of recombinant Cas9 WT , nCas9 D10A , and LbCas12a-ultra The pET-LbCas12a-ultra vector was constructed using the DNA sequence encoding LbCas12a-ultra (18), which was cloned into the pET28a+ vector (Addgene, #69864-3). pETCas9-HN was derived from p3s-Cas9-HN (Addgene, #104171, RRID: Addgene_104171). .. These vectors were transformed into the C3013 strain (NEB, #C3013I), and recombinant Cas9 WT or LbCas12a-ultra protein was expressed at 25°C for 4 h by addition of 0.5 mM IPTG, followed by purification using Ni-NTA agarose beads (Qiagen, #30210).

    Plasmid Preparation:

    Article Title: Combining Multiplexed CRISPR/Cas9-Nickase and PARP Inhibitors Efficiently and Precisely Targets Cancer Cells
    Article Snippet: .. Purification of recombinant Cas9 WT , nCas9 D10A , and LbCas12a-ultra The pET-LbCas12a-ultra vector was constructed using the DNA sequence encoding LbCas12a-ultra (18), which was cloned into the pET28a+ vector (Addgene, #69864-3). pETCas9-HN was derived from p3s-Cas9-HN (Addgene, #104171, RRID: Addgene_104171). .. These vectors were transformed into the C3013 strain (NEB, #C3013I), and recombinant Cas9 WT or LbCas12a-ultra protein was expressed at 25°C for 4 h by addition of 0.5 mM IPTG, followed by purification using Ni-NTA agarose beads (Qiagen, #30210).

    Article Title: Highly parallel profiling of the activities and specificities of Cas12a variants in human cells
    Article Snippet: .. AsCas12a (Addgene, #69982), LbCas12a (Addgene, #69988), FnCas12a (Addgene, #69976), Lb2Cas12a (Addgene, #69983) human expression plasmids were purchased from the non-profit plasmid repository Addgene. .. To prepare the lentiviral vectors, we removed the Cas9-encoding sequence from the lentiCas9-Blast plasmid (Addgene, #52962), and cloned Cas12a-encoding sequences between XbaI and BamHI.

    Construct:

    Article Title: Combining Multiplexed CRISPR/Cas9-Nickase and PARP Inhibitors Efficiently and Precisely Targets Cancer Cells
    Article Snippet: .. Purification of recombinant Cas9 WT , nCas9 D10A , and LbCas12a-ultra The pET-LbCas12a-ultra vector was constructed using the DNA sequence encoding LbCas12a-ultra (18), which was cloned into the pET28a+ vector (Addgene, #69864-3). pETCas9-HN was derived from p3s-Cas9-HN (Addgene, #104171, RRID: Addgene_104171). .. These vectors were transformed into the C3013 strain (NEB, #C3013I), and recombinant Cas9 WT or LbCas12a-ultra protein was expressed at 25°C for 4 h by addition of 0.5 mM IPTG, followed by purification using Ni-NTA agarose beads (Qiagen, #30210).

    Article Title: Highly efficient and specific regulation of gene expression using enhanced CRISPR-Cas12f system
    Article Snippet: Methods Design and cloning of various transcriptional activator and guide RNA expression vectors Cytomegalovirus (CMV) promoter-based expression vectors for dCWCas12f(D354A)-VPR, dLbCas12a(D832A)-VPR, and dSpCas9m4(D10A/D839A/H840A/N863A)-VPR were used to facilitate targeted gene expression in human-derived cell line (HEK293FT, Invitrogen (R70007)). .. LbCas12a(D832A)VPR (Addgene no. 104567) and SpCas9m4(D10A/D839A/H840A/N863A)-VPR (Addgene no. 63798) were constructed and used for experimental purposes. ..

    Sequencing:

    Article Title: Combining Multiplexed CRISPR/Cas9-Nickase and PARP Inhibitors Efficiently and Precisely Targets Cancer Cells
    Article Snippet: .. Purification of recombinant Cas9 WT , nCas9 D10A , and LbCas12a-ultra The pET-LbCas12a-ultra vector was constructed using the DNA sequence encoding LbCas12a-ultra (18), which was cloned into the pET28a+ vector (Addgene, #69864-3). pETCas9-HN was derived from p3s-Cas9-HN (Addgene, #104171, RRID: Addgene_104171). .. These vectors were transformed into the C3013 strain (NEB, #C3013I), and recombinant Cas9 WT or LbCas12a-ultra protein was expressed at 25°C for 4 h by addition of 0.5 mM IPTG, followed by purification using Ni-NTA agarose beads (Qiagen, #30210).

    Clone Assay:

    Article Title: Combining Multiplexed CRISPR/Cas9-Nickase and PARP Inhibitors Efficiently and Precisely Targets Cancer Cells
    Article Snippet: .. Purification of recombinant Cas9 WT , nCas9 D10A , and LbCas12a-ultra The pET-LbCas12a-ultra vector was constructed using the DNA sequence encoding LbCas12a-ultra (18), which was cloned into the pET28a+ vector (Addgene, #69864-3). pETCas9-HN was derived from p3s-Cas9-HN (Addgene, #104171, RRID: Addgene_104171). .. These vectors were transformed into the C3013 strain (NEB, #C3013I), and recombinant Cas9 WT or LbCas12a-ultra protein was expressed at 25°C for 4 h by addition of 0.5 mM IPTG, followed by purification using Ni-NTA agarose beads (Qiagen, #30210).

    Derivative Assay:

    Article Title: Combining Multiplexed CRISPR/Cas9-Nickase and PARP Inhibitors Efficiently and Precisely Targets Cancer Cells
    Article Snippet: .. Purification of recombinant Cas9 WT , nCas9 D10A , and LbCas12a-ultra The pET-LbCas12a-ultra vector was constructed using the DNA sequence encoding LbCas12a-ultra (18), which was cloned into the pET28a+ vector (Addgene, #69864-3). pETCas9-HN was derived from p3s-Cas9-HN (Addgene, #104171, RRID: Addgene_104171). .. These vectors were transformed into the C3013 strain (NEB, #C3013I), and recombinant Cas9 WT or LbCas12a-ultra protein was expressed at 25°C for 4 h by addition of 0.5 mM IPTG, followed by purification using Ni-NTA agarose beads (Qiagen, #30210).

    Cloning:

    Article Title: Hs1Cas12a and Ev1Cas12a confer efficient genome editing in plants
    Article Snippet: Cas12a entry clones and crRNA entry clones were assembled with the destination vector pYPQ203 (Addgene #86207, ) by Gateway LR reactions to generate the final T-DNA vectors ( ). .. LbCas12a and Mb2Cas12a were used as controls. pYPQ141-ZmUbi-RZ-Lb (Addgene #86197, ) was used for crRNA cloning of LbCas12a. ..



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    DNA blocker mediated crRNA scaffold remodeling regulates Cas12a trans - and cis- cleavage activities. ( A ) Schematic of length-dependent scaffold occlusion at the 5′ end for regulating Cas12a cis- and trans-cleavage activities. ( B and C ) Trans -cleavage fluorescence signals (bar plot) and cleavage efficiencies (line plot with markers) after 1 h for blockers introduced from the 5′ end (B) or 3′ end (C) of the scaffold. ( D and E ) Cis -cleavage fluorescence signals (bar plot) and cleavage efficiencies (line plot with markers) after 1 h for blockers introduced from the 5′ end (D) or 3′ end (E) of the scaffold. ( F ) Domain structure of <t>LbCas12a</t> (PDB 5ID6). ( G ) Molecular docking simulation of LbCas12a with conventional crRNA and ssDNA activator. ( H ) Molecular docking simulation of LbCas12a with scaffold-closed crRNA and ssDNA activator. Each experiment was repeated three times.
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    DNA blocker mediated crRNA scaffold remodeling regulates Cas12a trans - and cis- cleavage activities. ( A ) Schematic of length-dependent scaffold occlusion at the 5′ end for regulating Cas12a cis- and trans-cleavage activities. ( B and C ) Trans -cleavage fluorescence signals (bar plot) and cleavage efficiencies (line plot with markers) after 1 h for blockers introduced from the 5′ end (B) or 3′ end (C) of the scaffold. ( D and E ) Cis -cleavage fluorescence signals (bar plot) and cleavage efficiencies (line plot with markers) after 1 h for blockers introduced from the 5′ end (D) or 3′ end (E) of the scaffold. ( F ) Domain structure of <t>LbCas12a</t> (PDB 5ID6). ( G ) Molecular docking simulation of LbCas12a with conventional crRNA and ssDNA activator. ( H ) Molecular docking simulation of LbCas12a with scaffold-closed crRNA and ssDNA activator. Each experiment was repeated three times.
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    Image Search Results


    DNA blocker mediated crRNA scaffold remodeling regulates Cas12a trans - and cis- cleavage activities. ( A ) Schematic of length-dependent scaffold occlusion at the 5′ end for regulating Cas12a cis- and trans-cleavage activities. ( B and C ) Trans -cleavage fluorescence signals (bar plot) and cleavage efficiencies (line plot with markers) after 1 h for blockers introduced from the 5′ end (B) or 3′ end (C) of the scaffold. ( D and E ) Cis -cleavage fluorescence signals (bar plot) and cleavage efficiencies (line plot with markers) after 1 h for blockers introduced from the 5′ end (D) or 3′ end (E) of the scaffold. ( F ) Domain structure of LbCas12a (PDB 5ID6). ( G ) Molecular docking simulation of LbCas12a with conventional crRNA and ssDNA activator. ( H ) Molecular docking simulation of LbCas12a with scaffold-closed crRNA and ssDNA activator. Each experiment was repeated three times.

    Journal: Nucleic Acids Research

    Article Title: crRNA scaffold remodeling controls CRISPR–Cas12a activity for enhanced performance

    doi: 10.1093/nar/gkag167

    Figure Lengend Snippet: DNA blocker mediated crRNA scaffold remodeling regulates Cas12a trans - and cis- cleavage activities. ( A ) Schematic of length-dependent scaffold occlusion at the 5′ end for regulating Cas12a cis- and trans-cleavage activities. ( B and C ) Trans -cleavage fluorescence signals (bar plot) and cleavage efficiencies (line plot with markers) after 1 h for blockers introduced from the 5′ end (B) or 3′ end (C) of the scaffold. ( D and E ) Cis -cleavage fluorescence signals (bar plot) and cleavage efficiencies (line plot with markers) after 1 h for blockers introduced from the 5′ end (D) or 3′ end (E) of the scaffold. ( F ) Domain structure of LbCas12a (PDB 5ID6). ( G ) Molecular docking simulation of LbCas12a with conventional crRNA and ssDNA activator. ( H ) Molecular docking simulation of LbCas12a with scaffold-closed crRNA and ssDNA activator. Each experiment was repeated three times.

    Article Snippet: All reactions were assembled in a 20 μl volume containing the crRNA/blocker duplex (50 nM/75 nM), LbCas12a (50 nM), and NEBuffer r2.1 (1×), with TE Mg 2+ buffer (12.5 mM) added to reach the final volume.

    Techniques: Fluorescence