Nls sacas9 nls 3xha bghpa

Manufactured by Addgene

NLS-SaCas9-NLS-3xHA-bGHpA is a plasmid that encodes the Staphylococcus aureus Cas9 (SaCas9) protein with a nuclear localization signal (NLS) and a 3xHA epitope tag. The SaCas9 protein is a smaller version of the commonly used Streptococcus pyogenes Cas9 and can be used for CRISPR-Cas9 genome editing applications.

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Lab products found in correlation

Px601, by Addgene (1 mentions) Pspcas9 bb 2a gfp px458, by Addgene (1 mentions) U6 bsai sgrna, by Addgene (1 mentions)

2 protocols using nls sacas9 nls 3xha bghpa

CRISPR-Cas9 Plasmid Construction

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sgRNA sequences were designed using online software provided by Benchling (See next section for sequences). Plasmids: pX601-AAV-CMV: NLS-SaCas9-NLS-3xHA-bGHpA; U6: BsaI-sgRNA and pX601-GFP were obtained from Addgene. The paired gRNAs were synthesized and annealed according to standard protocols. Once annealed, the gRNAs were cloned into the vector by using BsaI restriction site.

Sun H., Hodgkinson C.P., Pratt R.E, & Dzau V.J. (2021). Crispr-Cas9 deletion of hepatic Angiotensinogen gene results in sustained control of Hypertension: A Potential for Cure?. Hypertension (Dallas, Tex. : 1979), 77(6), 1990-2000.

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Expression and Purification of CRISPR Proteins

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The gene sequences of SaCas9, SpCas9, AsCas12a, and LbCas12a proteins were derived from pX601-AAV-CMV::NLS-SaCas9-NLS-3xHA-bGHpA;U6::BsaI-sgRNA (Addgene plasmid # 61591), pSpCas9(BB)-2A-GFP (PX458) (Addgene plasmid # 48138), pY010 (pcDNA3.1-hAsCpf1) (Addgene plasmid # 69982), and pY016 (pcDNA3.1-hLbCpf1) (Addgene plasmid # 69988), respectively, which were gifts from Feng Zhang [33] (link)[34] (link)[35] (link). By comparing the fragments of the expression vector and four target genes, we selected SalI and XbaI as the insertion sites between the multiple clone site of the pET28b prokaryotic expression vector. Then primers were designed for the gene sequences of SaCas9, SpCas9, AsCas12a and LbCas12a. The forward primer 5' was added with SalI digestion site sequence, and the reverse primer

Xiao P., Bai R., Zhang T., Wu R., Chen L., Hou Y., Shen B., Niu Y., Li S., Ji W, & Chen Y. (2021). Extensive humoral immune response to AAVs and Cas proteins in nonhuman primates. Science bulletin, 66(20).

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