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Addgene inc bbsi site
Bbsi Site, supplied by Addgene inc, used in various techniques. Bioz Stars score: 96/100, based on 4065 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/bbsi+site/pSpCas9(BB)-2A-GFP+(PX458)+(Plasmid+%2348138)/bio_rxiv__64898__2026__01__22__701170-240-27-29
Average 96 stars, based on 4065 article reviews

bbsi site - by Bioz Stars, 2026-09

96/100 stars

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Clone Assay:

Article Title: A missing enzyme-rescue metabolite as cause of a rare skeletal dysplasia
Article Snippet: .. Complementary sgRNA oligonucleotides were subsequently annealed, phosphorylated and cloned into the BbsI site of dephosphorylated pX459 pSpCas9(BB)-2A-Puro vector (Addgene; #62988). ..

Article Title: Conformation-specific Antibody Deciphers K27-linked Ubiquitination in Chaperone-Mediated Proteostasis
Article Snippet: Human TRIM11 (NM_145214.3) and TRIM65 (NM_173547.4) was obtained from MiaoLing Plasmid and cloned into the BamHI / XhoI sites of pGEX-6p-1 for bacterial expression. .. For CRISPR-Cas9 genome knockout, double strands of single guide RNA targeting hDNAJB1 (5’-CACCGCGGGTCGCTGAGCACGTCGT-3’, and 5’-AAACACGACGTGCTCAGCGACCCGC-3’) were syntheses by Sangon Biotech and cloned into the BbsI site of PX459 backbone (Addgene #62988; obtained from MiaoLing Plasmid), termed PX459-sgDNAJB1#2. ..

Article Title: Cell-type-specific dysregulation of gene expression due to Chd8 haploinsufficiency during mouse cortical development
Article Snippet: .. The sgRNA linker sequences were cloned into the BbsI site of plasmid pX335-U6-Chimeric_BB-CBh-hSpCas9n(D10A) (Addgene, #42335, RRID: Addgene_42335) to generate a chimeric sequence of sgRNA-trRNA downstream of the T7 promoter. .. The plasmid containing the indicated sgRNA target sequences were individually PCR amplified and transcribed in vitro using the MEGAshortscript T7 kit (ThermoFisher Scientific, AM1354) and purified using the MEGAclear Transcription Clean-Up kit (ThermoFisher Scientific, AM1908).

CRISPR:

Knock-Out:

Plasmid Preparation:

Article Title: CRISPR-mediated conditional mutagenesis of Smad1/5/8 reveals BMP/GDF signaling restricts postnatal bone overgrowth
Article Snippet: gRNAs were designed using CHOPCHOP and synthesized as forward and reserve oligos by Integrated DNA Technologies (IA, USA). .. To validate and test the cutting efficiency of each gRNA, complementary oligos were ligated and cloned into px458 plasmid directly upstream of a gRNA scaffold at the BbsI site (Addgene plasmid #48138). .. Each plasmid was transfected into immortalized embryonic mouse fibroblasts (iMEFs) (ATCC, CRL-2907) using TransitX2 (Mirus, MIR 6004) in triplicate. iMEFs were incubated at 37°C for 72 hours post transfection in DMEM (ATCC, 30-2002) supplemented with 10% FBS (Omega Scientific, NC0471611) and 1% Pen/Strep (ThermoFisher, 15140122).

Article Title: Genetic Analysis of Choroideremia-Related Rab Escort Proteins
Article Snippet: .. We designed three single-guide RNAs (sgRNAs) and annealed their corresponding oligonucleotides, and then, we inserted them into the BbsI site of the pX330-U6Chimeric_BB-CBh-hSpCas9 vector (Addgene plasmid #42230, Watertown, MA, USA) [56]. .. T7 promoters were added to the sgRNA templates via PCR, and the sgRNAs were synthesized using the MEGAshortscriptTM T7 Transcription Kit (Invitrogen, AM1354, Waltham, MA, USA).

Sequencing:

other:

Article Title: Cell-type-specific dysregulation of gene expression due to Chd8 haploinsufficiency during mouse cortical development.
Article Snippet: 124 The plasmid containing the indicated sgRNA target sequences were individually PCR amplified and transcribed in vitro using the MEGAshortscript T7 kit (ThermoFisher Scientific, AM1354) and purified using the MEGAclear Transcription Clean-Up kit (ThermoFisher Scientific, AM1908).

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