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Addgene inc spcas9
Spcas9, supplied by Addgene inc, used in various techniques. Bioz Stars score: 96/100, based on 369 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/cas9+spcas9/SP-Cas9+(Plasmid+%2362731)/us12590323-392-17-24
Average 96 stars, based on 369 article reviews
spcas9 - by Bioz Stars, 2026-09
96/100 stars

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Related Articles

Sequencing:

Article Title: A novel Cas9 fusion protein promotes targeted genome editing with reduced mutational burden in primary human cells.
Article Snippet: .. The Streptococcus pyogenes Cas9 (spCas9) coding sequence was amplified via PCR from lentiCRISPR 1516 (Addgene 49525) and cloned into a plasmid containing two nuclear localization signals (NLS) both at N- and C-terminus of the Cas9 and a flexible (G4S) linker at the C-terminus of the Cas9. .. The C-terminus fusions were generated by cleaving the resulting spCas9 expression plasmid with AvrII (NEB) and PmeI (NEB).

Article Title: A novel Cas9 fusion protein promotes targeted genome editing with reduced mutational burden in primary human cells
Article Snippet: .. The Streptococcus pyogenes Cas9 (spCas9) coding sequence was amplified via PCR from lentiCRISPR_1516 (Addgene 49525) and cloned into a plasmid containing two nuclear localization signals (NLS) both at N- and C-terminus of the Cas9 and a flexible (G4S) linker at the C-terminus of the Cas9. .. The C-terminus fusions were generated by cleaving the resulting spCas9 expression plasmid with AvrII (NEB) and PmeI (NEB).

Amplification:

Article Title: A novel Cas9 fusion protein promotes targeted genome editing with reduced mutational burden in primary human cells.
Article Snippet: .. The Streptococcus pyogenes Cas9 (spCas9) coding sequence was amplified via PCR from lentiCRISPR 1516 (Addgene 49525) and cloned into a plasmid containing two nuclear localization signals (NLS) both at N- and C-terminus of the Cas9 and a flexible (G4S) linker at the C-terminus of the Cas9. .. The C-terminus fusions were generated by cleaving the resulting spCas9 expression plasmid with AvrII (NEB) and PmeI (NEB).

Article Title: A novel Cas9 fusion protein promotes targeted genome editing with reduced mutational burden in primary human cells
Article Snippet: .. The Streptococcus pyogenes Cas9 (spCas9) coding sequence was amplified via PCR from lentiCRISPR_1516 (Addgene 49525) and cloned into a plasmid containing two nuclear localization signals (NLS) both at N- and C-terminus of the Cas9 and a flexible (G4S) linker at the C-terminus of the Cas9. .. The C-terminus fusions were generated by cleaving the resulting spCas9 expression plasmid with AvrII (NEB) and PmeI (NEB).

Polymerase Chain Reaction:

Article Title: A novel Cas9 fusion protein promotes targeted genome editing with reduced mutational burden in primary human cells.
Article Snippet: .. The Streptococcus pyogenes Cas9 (spCas9) coding sequence was amplified via PCR from lentiCRISPR 1516 (Addgene 49525) and cloned into a plasmid containing two nuclear localization signals (NLS) both at N- and C-terminus of the Cas9 and a flexible (G4S) linker at the C-terminus of the Cas9. .. The C-terminus fusions were generated by cleaving the resulting spCas9 expression plasmid with AvrII (NEB) and PmeI (NEB).

Article Title: A novel Cas9 fusion protein promotes targeted genome editing with reduced mutational burden in primary human cells
Article Snippet: .. The Streptococcus pyogenes Cas9 (spCas9) coding sequence was amplified via PCR from lentiCRISPR_1516 (Addgene 49525) and cloned into a plasmid containing two nuclear localization signals (NLS) both at N- and C-terminus of the Cas9 and a flexible (G4S) linker at the C-terminus of the Cas9. .. The C-terminus fusions were generated by cleaving the resulting spCas9 expression plasmid with AvrII (NEB) and PmeI (NEB).

Clone Assay:

Article Title: A novel Cas9 fusion protein promotes targeted genome editing with reduced mutational burden in primary human cells.
Article Snippet: .. The Streptococcus pyogenes Cas9 (spCas9) coding sequence was amplified via PCR from lentiCRISPR 1516 (Addgene 49525) and cloned into a plasmid containing two nuclear localization signals (NLS) both at N- and C-terminus of the Cas9 and a flexible (G4S) linker at the C-terminus of the Cas9. .. The C-terminus fusions were generated by cleaving the resulting spCas9 expression plasmid with AvrII (NEB) and PmeI (NEB).

Article Title: A novel Cas9 fusion protein promotes targeted genome editing with reduced mutational burden in primary human cells
Article Snippet: .. The Streptococcus pyogenes Cas9 (spCas9) coding sequence was amplified via PCR from lentiCRISPR_1516 (Addgene 49525) and cloned into a plasmid containing two nuclear localization signals (NLS) both at N- and C-terminus of the Cas9 and a flexible (G4S) linker at the C-terminus of the Cas9. .. The C-terminus fusions were generated by cleaving the resulting spCas9 expression plasmid with AvrII (NEB) and PmeI (NEB).

Plasmid Preparation:

Article Title: A novel Cas9 fusion protein promotes targeted genome editing with reduced mutational burden in primary human cells.
Article Snippet: .. The Streptococcus pyogenes Cas9 (spCas9) coding sequence was amplified via PCR from lentiCRISPR 1516 (Addgene 49525) and cloned into a plasmid containing two nuclear localization signals (NLS) both at N- and C-terminus of the Cas9 and a flexible (G4S) linker at the C-terminus of the Cas9. .. The C-terminus fusions were generated by cleaving the resulting spCas9 expression plasmid with AvrII (NEB) and PmeI (NEB).

Article Title: Exosomal PD-L1 promotes tumor growth through immune escape in non-small cell lung cancer.
Article Snippet: Constructs for the suppression of human PD-L1 (hPD-L1) and overexpression of murine PD-L1 (mPD-L1) For the generation of lentiviral-based human PD-L1 via CRISPR/Cas9, a single-guide RNA (sgRNA) specifically recognizing and potentially destroying the open reading frame of the hPD-L1 gene was selected using a web-based tool, Benchling (https://benchling.com/). .. To construct an expression vector for Streptococcus pyogenes Cas9 (SpCas9) and an sgRNA specific for the hPD-L1 gene, a lentiviral vector (lentiCRISPR v2, Addgene #52961) was obtained from Addgene (Cambridge, MA, USA), and annealed oligomers (5′-CACCGTCTTTATATTCATGA CCTAC-3′ and 5′-AAACGTAGGTCATGAATATAAA GAC-3′) was subcloned using the BsmB1 sites, as previously described26. .. For Sanger sequencing (Macrogen, Inc., Seoul, Korea), the following primers were used for polymerase chain reaction (PCR) analyses: 5′-CAGTTAG AACCACCAAGTCCCA-3′ and 5′-AGGATCTTGGCC TTGTTGAAA-3′ (464 bp for the wild-type PD-L1 gene).

Article Title: Nonspecific interactions between SpCas9 and dsDNA sites located downstream of the PAM mediate facilitated diffusion to accelerate target search
Article Snippet: .. S. pyogenes Cas9 (SpCas9) were expressed in Escherichia coli strain BL21 (DE3) using the expression plasmid pMJ806 (Addgene plasmid # 39312) (Jinek et al., 2012) and purified as previously described 1. ..

Article Title: A novel Cas9 fusion protein promotes targeted genome editing with reduced mutational burden in primary human cells
Article Snippet: .. The Streptococcus pyogenes Cas9 (spCas9) coding sequence was amplified via PCR from lentiCRISPR_1516 (Addgene 49525) and cloned into a plasmid containing two nuclear localization signals (NLS) both at N- and C-terminus of the Cas9 and a flexible (G4S) linker at the C-terminus of the Cas9. .. The C-terminus fusions were generated by cleaving the resulting spCas9 expression plasmid with AvrII (NEB) and PmeI (NEB).

Article Title: Systematic in vitro specificity profiling reveals nicking defects in natural and engineered CRISPR–Cas9 variants
Article Snippet: Expression plasmids for SpCas9 and high-fidelity variants were purchased from Addgene. .. Streptococcus pyogenes Cas9 (SpCas9) (pMJ806) was expressed using expression plasmid pEC-K-MBP, and SpCas9-HF1 (pJSC111) and HypaCas9 (pJSC173) were expressed using expression plasmid pCT10. pMJ806, pJSC111, pJSC173 were gifts from Jennifer Doudna and/or Keith Joung (Addgene plasmid #39312; http://n2t.net/addgene:39312; RRID:Addgene_39312; Addgene plasmid # 101209; http://n2t.net/addgene:101209; RRID:Addgene_101209; Addgene plasmid #101218; http://n2t.net/addgene:101218; RRID:Addgene_101218). .. The gene sequence for S. aureus Cas9 (SaCas9) was synthesized as Escherichia coli codon-optimized gBlocks (purchased from Integrated DNA Technologies, IDT).

Article Title: High-throughput in vitro specificity profiling of natural and high-fidelity CRISPR-Cas9 variants
Article Snippet: Expression plasmids for SpCas9 and high-fidelity variants were purchased from Addgene. .. Streptococcus pyogenes Cas9 (SpCas9) (pMJ806) was expressed using expression plasmid pEC-K-MBP, and SpCas9-HF1 (pJSC111) and HypaCas9 (pJSC173) were expressed using expression plasmid pCT10. pMJ806, pJSC111, pJSC173 were gifts from Jennifer Doudna and/or Keith Joung (Addgene plasmid # 39312; http://n2t.net/addgene:39312 ; RRID:Addgene_39312; Addgene plasmid # 101209; http://n2t.net/addgene:101209 ; RRID:Addgene_101209; Addgene plasmid # 101218; http://n2t.net/addgene:101218 ; RRID:Addgene_101218). .. The gene sequence for Staphylococcus aureus Cas9 (SaCas9) was synthesized as Escherichia coli codon-optimized gBlocks (purchased from Integrated DNA Technologies, IDT).

Construct:

Article Title: Exosomal PD-L1 promotes tumor growth through immune escape in non-small cell lung cancer.
Article Snippet: Constructs for the suppression of human PD-L1 (hPD-L1) and overexpression of murine PD-L1 (mPD-L1) For the generation of lentiviral-based human PD-L1 via CRISPR/Cas9, a single-guide RNA (sgRNA) specifically recognizing and potentially destroying the open reading frame of the hPD-L1 gene was selected using a web-based tool, Benchling (https://benchling.com/). .. To construct an expression vector for Streptococcus pyogenes Cas9 (SpCas9) and an sgRNA specific for the hPD-L1 gene, a lentiviral vector (lentiCRISPR v2, Addgene #52961) was obtained from Addgene (Cambridge, MA, USA), and annealed oligomers (5′-CACCGTCTTTATATTCATGA CCTAC-3′ and 5′-AAACGTAGGTCATGAATATAAA GAC-3′) was subcloned using the BsmB1 sites, as previously described26. .. For Sanger sequencing (Macrogen, Inc., Seoul, Korea), the following primers were used for polymerase chain reaction (PCR) analyses: 5′-CAGTTAG AACCACCAAGTCCCA-3′ and 5′-AGGATCTTGGCC TTGTTGAAA-3′ (464 bp for the wild-type PD-L1 gene).

Expressing:

Article Title: Exosomal PD-L1 promotes tumor growth through immune escape in non-small cell lung cancer.
Article Snippet: Constructs for the suppression of human PD-L1 (hPD-L1) and overexpression of murine PD-L1 (mPD-L1) For the generation of lentiviral-based human PD-L1 via CRISPR/Cas9, a single-guide RNA (sgRNA) specifically recognizing and potentially destroying the open reading frame of the hPD-L1 gene was selected using a web-based tool, Benchling (https://benchling.com/). .. To construct an expression vector for Streptococcus pyogenes Cas9 (SpCas9) and an sgRNA specific for the hPD-L1 gene, a lentiviral vector (lentiCRISPR v2, Addgene #52961) was obtained from Addgene (Cambridge, MA, USA), and annealed oligomers (5′-CACCGTCTTTATATTCATGA CCTAC-3′ and 5′-AAACGTAGGTCATGAATATAAA GAC-3′) was subcloned using the BsmB1 sites, as previously described26. .. For Sanger sequencing (Macrogen, Inc., Seoul, Korea), the following primers were used for polymerase chain reaction (PCR) analyses: 5′-CAGTTAG AACCACCAAGTCCCA-3′ and 5′-AGGATCTTGGCC TTGTTGAAA-3′ (464 bp for the wild-type PD-L1 gene).

Article Title: Nonspecific interactions between SpCas9 and dsDNA sites located downstream of the PAM mediate facilitated diffusion to accelerate target search
Article Snippet: .. S. pyogenes Cas9 (SpCas9) were expressed in Escherichia coli strain BL21 (DE3) using the expression plasmid pMJ806 (Addgene plasmid # 39312) (Jinek et al., 2012) and purified as previously described 1. ..

Article Title: Systematic in vitro specificity profiling reveals nicking defects in natural and engineered CRISPR–Cas9 variants
Article Snippet: Expression plasmids for SpCas9 and high-fidelity variants were purchased from Addgene. .. Streptococcus pyogenes Cas9 (SpCas9) (pMJ806) was expressed using expression plasmid pEC-K-MBP, and SpCas9-HF1 (pJSC111) and HypaCas9 (pJSC173) were expressed using expression plasmid pCT10. pMJ806, pJSC111, pJSC173 were gifts from Jennifer Doudna and/or Keith Joung (Addgene plasmid #39312; http://n2t.net/addgene:39312; RRID:Addgene_39312; Addgene plasmid # 101209; http://n2t.net/addgene:101209; RRID:Addgene_101209; Addgene plasmid #101218; http://n2t.net/addgene:101218; RRID:Addgene_101218). .. The gene sequence for S. aureus Cas9 (SaCas9) was synthesized as Escherichia coli codon-optimized gBlocks (purchased from Integrated DNA Technologies, IDT).

Article Title: High-throughput in vitro specificity profiling of natural and high-fidelity CRISPR-Cas9 variants
Article Snippet: Expression plasmids for SpCas9 and high-fidelity variants were purchased from Addgene. .. Streptococcus pyogenes Cas9 (SpCas9) (pMJ806) was expressed using expression plasmid pEC-K-MBP, and SpCas9-HF1 (pJSC111) and HypaCas9 (pJSC173) were expressed using expression plasmid pCT10. pMJ806, pJSC111, pJSC173 were gifts from Jennifer Doudna and/or Keith Joung (Addgene plasmid # 39312; http://n2t.net/addgene:39312 ; RRID:Addgene_39312; Addgene plasmid # 101209; http://n2t.net/addgene:101209 ; RRID:Addgene_101209; Addgene plasmid # 101218; http://n2t.net/addgene:101218 ; RRID:Addgene_101218). .. The gene sequence for Staphylococcus aureus Cas9 (SaCas9) was synthesized as Escherichia coli codon-optimized gBlocks (purchased from Integrated DNA Technologies, IDT).

Purification:

Article Title: Nonspecific interactions between SpCas9 and dsDNA sites located downstream of the PAM mediate facilitated diffusion to accelerate target search
Article Snippet: .. S. pyogenes Cas9 (SpCas9) were expressed in Escherichia coli strain BL21 (DE3) using the expression plasmid pMJ806 (Addgene plasmid # 39312) (Jinek et al., 2012) and purified as previously described 1. ..

Modification:

Article Title: Loss of Emp2 compromises cardiogenic differentiation in mouse embryonic stem cells.
Article Snippet: Isolated mouse embryonic stem cells (mESCs) retain the capacities to self-renew limitlessly and to give rise to all tissues of an adult mouse.. A precise understanding of the relationships, mechanisms of action and functions of novel genes involved in mESCs differentiation is crucial to expand our knowledge of vertebrate development.. The epithelial membrane protein 2 (EMP2) is a membrane-spanning protein found in epithelial and endothelial cell-cell junctions that has been implicated in the regulation of cell proliferation and migration in normal and tumor tissues.

Derivative Assay:

Article Title: High-efficiency and multilocus targeted integration in CHO cells using CRISPR-mediated donor nicking and DNA repair inhibitors.
Article Snippet: .. All Cas9 nuclease variants used in this study were derived from Streptococcus pyogenes Cas9 (spCas9) and expressed from plasmids. hCas9 (purchased from Addgene, #41815) and hCas9_D10A (purchased from Addgene, #41816) were used to make hCas9_H840A and hCas9_D10A_H840A (expresses nuclease‐dead Cas9 [dCas9]), respectively, through site‐directed mutagenesis to incorporate H840A substitutions. ..

Mutagenesis:

Article Title: High-efficiency and multilocus targeted integration in CHO cells using CRISPR-mediated donor nicking and DNA repair inhibitors.
Article Snippet: .. All Cas9 nuclease variants used in this study were derived from Streptococcus pyogenes Cas9 (spCas9) and expressed from plasmids. hCas9 (purchased from Addgene, #41815) and hCas9_D10A (purchased from Addgene, #41816) were used to make hCas9_H840A and hCas9_D10A_H840A (expresses nuclease‐dead Cas9 [dCas9]), respectively, through site‐directed mutagenesis to incorporate H840A substitutions. ..



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