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crispr/cas9 constructs  (Addgene inc)


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    Addgene inc crispr/cas9 constructs
    Crispr/Cas9 Constructs, 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/crispr/cas9 constructs/product/Addgene inc
    Average 90 stars, based on 1 article reviews
    crispr/cas9 constructs - by Bioz Stars, 2026-03
    90/100 stars

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    transfected construct (crispr/cas9)  (Benchling Inc)
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    ( A ) Outline of RNP CRISPR editing. The CTNS gene is located on chromosome 17p13.3 and consists of 12 exons, of which the first 2 are non-coding. Therefore, the guide RNA was targeted towards exon 3 to completely knockout the functional CTNS gene. <t>CRISPR-Cas9</t> ribonucleoproteins (crRNPs) were synthesized in vitro to knockout CTNS gene and delivered to immortalize RPTECs by nucleofection. These cells were expanded for molecular validation of gene editing and downstream functional assays. ( B ) The guide RNA target sequence and associated PAM are highlighted on the right. We performed Sanger sequencing and the TIDE output calculating percent indels from chromatograms is depicted in the bar-graph. ( C ) Validation of CTNS knockout (-/-) in immortalized RPTEC by using two primers targeting different exons – CTNS #1 targets between exon 2–3 and CTNS #2 targets between exon 9–10. We have shown that in the CRISPR/Cas9 CTNS -/- RPTEC there is a significant reduced Cystinosin RNA levels with both the primers. ( D ) Validation of CTNS -/- in immortalized RPTEC. Increased intracellular accumulation of cystine is shown by HPLC-MS/MS method in control and CTNS -/- RPTECs. Student’s t-test was used. Data are presented as mean ± SD. *p<0.05; **p<0.01; ***p<0.001.
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    ( A ) Outline of RNP CRISPR editing. The CTNS gene is located on chromosome 17p13.3 and consists of 12 exons, of which the first 2 are non-coding. Therefore, the guide RNA was targeted towards exon 3 to completely knockout the functional CTNS gene. CRISPR-Cas9 ribonucleoproteins (crRNPs) were synthesized in vitro to knockout CTNS gene and delivered to immortalize RPTECs by nucleofection. These cells were expanded for molecular validation of gene editing and downstream functional assays. ( B ) The guide RNA target sequence and associated PAM are highlighted on the right. We performed Sanger sequencing and the TIDE output calculating percent indels from chromatograms is depicted in the bar-graph. ( C ) Validation of CTNS knockout (-/-) in immortalized RPTEC by using two primers targeting different exons – CTNS #1 targets between exon 2–3 and CTNS #2 targets between exon 9–10. We have shown that in the CRISPR/Cas9 CTNS -/- RPTEC there is a significant reduced Cystinosin RNA levels with both the primers. ( D ) Validation of CTNS -/- in immortalized RPTEC. Increased intracellular accumulation of cystine is shown by HPLC-MS/MS method in control and CTNS -/- RPTECs. Student’s t-test was used. Data are presented as mean ± SD. *p<0.05; **p<0.01; ***p<0.001.

    Journal: eLife

    Article Title: Novel mechanism for tubular injury in nephropathic cystinosis

    doi: 10.7554/eLife.94169

    Figure Lengend Snippet: ( A ) Outline of RNP CRISPR editing. The CTNS gene is located on chromosome 17p13.3 and consists of 12 exons, of which the first 2 are non-coding. Therefore, the guide RNA was targeted towards exon 3 to completely knockout the functional CTNS gene. CRISPR-Cas9 ribonucleoproteins (crRNPs) were synthesized in vitro to knockout CTNS gene and delivered to immortalize RPTECs by nucleofection. These cells were expanded for molecular validation of gene editing and downstream functional assays. ( B ) The guide RNA target sequence and associated PAM are highlighted on the right. We performed Sanger sequencing and the TIDE output calculating percent indels from chromatograms is depicted in the bar-graph. ( C ) Validation of CTNS knockout (-/-) in immortalized RPTEC by using two primers targeting different exons – CTNS #1 targets between exon 2–3 and CTNS #2 targets between exon 9–10. We have shown that in the CRISPR/Cas9 CTNS -/- RPTEC there is a significant reduced Cystinosin RNA levels with both the primers. ( D ) Validation of CTNS -/- in immortalized RPTEC. Increased intracellular accumulation of cystine is shown by HPLC-MS/MS method in control and CTNS -/- RPTECs. Student’s t-test was used. Data are presented as mean ± SD. *p<0.05; **p<0.01; ***p<0.001.

    Article Snippet: Transfected Construct (CRISPR/cas9) ( Homo-sapiens ) , guide RNA, tracrRNA, and Cas9 , Benchling/Dharmacon , , .

    Techniques: CRISPR, Knock-Out, Functional Assay, Synthesized, In Vitro, Biomarker Discovery, Sequencing, Tandem Mass Spectroscopy, Control

    Journal: eLife

    Article Title: Novel mechanism for tubular injury in nephropathic cystinosis

    doi: 10.7554/eLife.94169

    Figure Lengend Snippet:

    Article Snippet: Transfected Construct (CRISPR/cas9) ( Homo-sapiens ) , guide RNA, tracrRNA, and Cas9 , Benchling/Dharmacon , , .

    Techniques: Transfection, Construct, CRISPR, Sequencing, Microarray, Isolation, Recombinant, Plasmid Preparation, Membrane