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Schematic for <t>CRISPR/Cas9</t> system into human AAVS1 locus. Top: Human AAVS1 locus. Single guide RNA (sgRNA) is indicated above the AAVS1 locus. Center: Donor DNA with 5’ homology arm (HA) and 3’HA, ∼820 bp on each side (dashed lines), splice acceptor sequence, T2A ribosome-skipping sequences, puromycin resistance sequence, Red Fluorescent Protein sequence (RFP), ancestral uricase sequence (AncUOX), and polyA signal. Bottom: Cas9 cleavage at the cut site allows donor DNA to be integrated via homology directed repair (HDR). After HDR the engineered AAVS1 locus is able to expresses Puromycin-resistance protein, RFP, and AncUOX.
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https://www.bioz.com/result/px458 plasmid/product/Addgene inc
Average 90 stars, based on 1 article reviews
px458 plasmid - by Bioz Stars, 2026-03
90/100 stars
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pspcas9(bb)-2a-gfp (px458)  (Addgene inc)
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Addgene inc pspcas9(bb)-2a-gfp (px458)
Schematic for <t>CRISPR/Cas9</t> system into human AAVS1 locus. Top: Human AAVS1 locus. Single guide RNA (sgRNA) is indicated above the AAVS1 locus. Center: Donor DNA with 5’ homology arm (HA) and 3’HA, ∼820 bp on each side (dashed lines), splice acceptor sequence, T2A ribosome-skipping sequences, puromycin resistance sequence, Red Fluorescent Protein sequence (RFP), ancestral uricase sequence (AncUOX), and polyA signal. Bottom: Cas9 cleavage at the cut site allows donor DNA to be integrated via homology directed repair (HDR). After HDR the engineered AAVS1 locus is able to expresses Puromycin-resistance protein, RFP, and AncUOX.
Pspcas9(Bb) 2a Gfp (Px458), supplied by Addgene inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/pspcas9(bb)-2a-gfp (px458)/product/Addgene inc
Average 90 stars, based on 1 article reviews
pspcas9(bb)-2a-gfp (px458) - by Bioz Stars, 2026-03
90/100 stars
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Schematic for CRISPR/Cas9 system into human AAVS1 locus. Top: Human AAVS1 locus. Single guide RNA (sgRNA) is indicated above the AAVS1 locus. Center: Donor DNA with 5’ homology arm (HA) and 3’HA, ∼820 bp on each side (dashed lines), splice acceptor sequence, T2A ribosome-skipping sequences, puromycin resistance sequence, Red Fluorescent Protein sequence (RFP), ancestral uricase sequence (AncUOX), and polyA signal. Bottom: Cas9 cleavage at the cut site allows donor DNA to be integrated via homology directed repair (HDR). After HDR the engineered AAVS1 locus is able to expresses Puromycin-resistance protein, RFP, and AncUOX.

Journal: Scientific Reports

Article Title: Genomic insertion of ancestral uricase into human liver cells to determine metabolic consequences of pseudogenization

doi: 10.1038/s41598-025-10551-8

Figure Lengend Snippet: Schematic for CRISPR/Cas9 system into human AAVS1 locus. Top: Human AAVS1 locus. Single guide RNA (sgRNA) is indicated above the AAVS1 locus. Center: Donor DNA with 5’ homology arm (HA) and 3’HA, ∼820 bp on each side (dashed lines), splice acceptor sequence, T2A ribosome-skipping sequences, puromycin resistance sequence, Red Fluorescent Protein sequence (RFP), ancestral uricase sequence (AncUOX), and polyA signal. Bottom: Cas9 cleavage at the cut site allows donor DNA to be integrated via homology directed repair (HDR). After HDR the engineered AAVS1 locus is able to expresses Puromycin-resistance protein, RFP, and AncUOX.

Article Snippet: The pX458 plasmid expressing Cas9 protein, gRNA and GFP were purchased from Addgene (#48138) and digested with Bbs I enzyme (New England Biolabs) .

Techniques: CRISPR, Sequencing