ppbu6 hdhfr yfcu Search Results


ppbu6 hdhfr yfcu cas9  (Addgene inc)
96
Addgene inc ppbu6 hdhfr yfcu cas9
( A ) Schematic of <t>pPbU6-hdhfr/yfcu-Cas9</t> with the homology regions (HR) of the repair template delivered on the same plasmid (one-plasmid approach) or on a PCR product (PCR-template approach) to generate the rap2/3 -3xHA parasite line. The effect on transfection efficiency was compared by both ( i ) substituting the P. yoelii U6 for the P. berghei U6 promoter (not significant) and ( ii ) different homology arm length (p = 0.0068). ( iii ) The relationship between homology arm length and transfection efficiency was examined by plotting homology arm length against the day parasitemia reached >0.5%. One or two-way ANOVAs were performed to compare transfection efficiencies from two biological replicates per condition. All parasitemia counts can be found in Table S2 . ( B ) The expression of RAP2/3-3xHA was confirmed by Western blot following transfection with the pPbU6-hdhfr/yfcu-Cas9 vector using ∼500 bp HR. Wild type parasites were used as a negative control for the HA tag and the anti-EXP1 was used as a loading control. Full-length Western blots are shown in Fig. S2 . ( C ) Growth assay of the PbCas9 line that constitutively expresses FLAG-Cas9 under the hsp70 promoter with the PbGIMO mother line used as a control. Error bars = SD for three biological replicates. ( D ) FLAG-Cas9 expression was confirmed using Western blot, where FLAG detects Cas9 and anti-EXP1 was used as loading control. PbGIMO was used as negative control for the FLAG tag. ( E ) Immunofluorescence assay demonstrates that FLAG-Cas9 is expressed and localised to the nucleus. FLAG detects Cas9 and 4′,6-diamidino-2-phenylindole (DAPI) stains the nucleus. BF = Bright-field. The PbGIMO mother line was used as a negative control. Scale bar = 5 µm. ( F ) Transfection efficiency of the pPbU6-hdhfr/yfcu rap2/3 -3xHA vector into the PbCas9 line was assessed using the one-plasmid versus the PCR-template approach. Error bars = SD for two biological replicates.
Ppbu6 Hdhfr Yfcu Cas9, supplied by Addgene inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/ppbu6+hdhfr+yfcu/SP-Cas9+(Plasmid+%2362731)/bio_rxiv__2024__04__20__590404-205-5-22
Average 96 stars, based on 1 article reviews
ppbu6 hdhfr yfcu cas9 - by Bioz Stars, 2026-09
96/100 stars
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ppbu6 hdhfr yfcu  (Valiant Co Ltd)
94
Valiant Co Ltd ppbu6 hdhfr yfcu
( A ) Schematic of <t>pPbU6-hdhfr/yfcu-Cas9</t> with the homology regions (HR) of the repair template delivered on the same plasmid (one-plasmid approach) or on a PCR product (PCR-template approach) to generate the rap2/3 -3xHA parasite line. The effect on transfection efficiency was compared by both ( i ) substituting the P. yoelii U6 for the P. berghei U6 promoter (not significant) and ( ii ) different homology arm length (p = 0.0068). ( iii ) The relationship between homology arm length and transfection efficiency was examined by plotting homology arm length against the day parasitemia reached >0.5%. One or two-way ANOVAs were performed to compare transfection efficiencies from two biological replicates per condition. All parasitemia counts can be found in Table S2 . ( B ) The expression of RAP2/3-3xHA was confirmed by Western blot following transfection with the pPbU6-hdhfr/yfcu-Cas9 vector using ∼500 bp HR. Wild type parasites were used as a negative control for the HA tag and the anti-EXP1 was used as a loading control. Full-length Western blots are shown in Fig. S2 . ( C ) Growth assay of the PbCas9 line that constitutively expresses FLAG-Cas9 under the hsp70 promoter with the PbGIMO mother line used as a control. Error bars = SD for three biological replicates. ( D ) FLAG-Cas9 expression was confirmed using Western blot, where FLAG detects Cas9 and anti-EXP1 was used as loading control. PbGIMO was used as negative control for the FLAG tag. ( E ) Immunofluorescence assay demonstrates that FLAG-Cas9 is expressed and localised to the nucleus. FLAG detects Cas9 and 4′,6-diamidino-2-phenylindole (DAPI) stains the nucleus. BF = Bright-field. The PbGIMO mother line was used as a negative control. Scale bar = 5 µm. ( F ) Transfection efficiency of the pPbU6-hdhfr/yfcu rap2/3 -3xHA vector into the PbCas9 line was assessed using the one-plasmid versus the PCR-template approach. Error bars = SD for two biological replicates.
Ppbu6 Hdhfr Yfcu, supplied by Valiant Co Ltd, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/ppbu6+hdhfr+yfcu/Pyrimethamine/pm39844455-167-5-15
Average 94 stars, based on 1 article reviews
ppbu6 hdhfr yfcu - by Bioz Stars, 2026-09
94/100 stars
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Image Search Results


( A ) Schematic of pPbU6-hdhfr/yfcu-Cas9 with the homology regions (HR) of the repair template delivered on the same plasmid (one-plasmid approach) or on a PCR product (PCR-template approach) to generate the rap2/3 -3xHA parasite line. The effect on transfection efficiency was compared by both ( i ) substituting the P. yoelii U6 for the P. berghei U6 promoter (not significant) and ( ii ) different homology arm length (p = 0.0068). ( iii ) The relationship between homology arm length and transfection efficiency was examined by plotting homology arm length against the day parasitemia reached >0.5%. One or two-way ANOVAs were performed to compare transfection efficiencies from two biological replicates per condition. All parasitemia counts can be found in Table S2 . ( B ) The expression of RAP2/3-3xHA was confirmed by Western blot following transfection with the pPbU6-hdhfr/yfcu-Cas9 vector using ∼500 bp HR. Wild type parasites were used as a negative control for the HA tag and the anti-EXP1 was used as a loading control. Full-length Western blots are shown in Fig. S2 . ( C ) Growth assay of the PbCas9 line that constitutively expresses FLAG-Cas9 under the hsp70 promoter with the PbGIMO mother line used as a control. Error bars = SD for three biological replicates. ( D ) FLAG-Cas9 expression was confirmed using Western blot, where FLAG detects Cas9 and anti-EXP1 was used as loading control. PbGIMO was used as negative control for the FLAG tag. ( E ) Immunofluorescence assay demonstrates that FLAG-Cas9 is expressed and localised to the nucleus. FLAG detects Cas9 and 4′,6-diamidino-2-phenylindole (DAPI) stains the nucleus. BF = Bright-field. The PbGIMO mother line was used as a negative control. Scale bar = 5 µm. ( F ) Transfection efficiency of the pPbU6-hdhfr/yfcu rap2/3 -3xHA vector into the PbCas9 line was assessed using the one-plasmid versus the PCR-template approach. Error bars = SD for two biological replicates.

Journal: bioRxiv

Article Title: A scalable CRISPR-Cas9 gene editing system facilitates CRISPR screens in the malaria parasite Plasmodium berghei

doi: 10.1101/2024.04.20.590404

Figure Lengend Snippet: ( A ) Schematic of pPbU6-hdhfr/yfcu-Cas9 with the homology regions (HR) of the repair template delivered on the same plasmid (one-plasmid approach) or on a PCR product (PCR-template approach) to generate the rap2/3 -3xHA parasite line. The effect on transfection efficiency was compared by both ( i ) substituting the P. yoelii U6 for the P. berghei U6 promoter (not significant) and ( ii ) different homology arm length (p = 0.0068). ( iii ) The relationship between homology arm length and transfection efficiency was examined by plotting homology arm length against the day parasitemia reached >0.5%. One or two-way ANOVAs were performed to compare transfection efficiencies from two biological replicates per condition. All parasitemia counts can be found in Table S2 . ( B ) The expression of RAP2/3-3xHA was confirmed by Western blot following transfection with the pPbU6-hdhfr/yfcu-Cas9 vector using ∼500 bp HR. Wild type parasites were used as a negative control for the HA tag and the anti-EXP1 was used as a loading control. Full-length Western blots are shown in Fig. S2 . ( C ) Growth assay of the PbCas9 line that constitutively expresses FLAG-Cas9 under the hsp70 promoter with the PbGIMO mother line used as a control. Error bars = SD for three biological replicates. ( D ) FLAG-Cas9 expression was confirmed using Western blot, where FLAG detects Cas9 and anti-EXP1 was used as loading control. PbGIMO was used as negative control for the FLAG tag. ( E ) Immunofluorescence assay demonstrates that FLAG-Cas9 is expressed and localised to the nucleus. FLAG detects Cas9 and 4′,6-diamidino-2-phenylindole (DAPI) stains the nucleus. BF = Bright-field. The PbGIMO mother line was used as a negative control. Scale bar = 5 µm. ( F ) Transfection efficiency of the pPbU6-hdhfr/yfcu rap2/3 -3xHA vector into the PbCas9 line was assessed using the one-plasmid versus the PCR-template approach. Error bars = SD for two biological replicates.

Article Snippet: The resultant vectors were named pPbU6-hdhfr/yfcu-Cas9 (addgene ID 216423, derived from pYCm and containing the coding sequence for spCas9 nuclease) and pPbU6-hdhfr/yfcu (Addgene #216422, derived from pYCs).

Techniques: Plasmid Preparation, Transfection, Expressing, Western Blot, Negative Control, Growth Assay, FLAG-tag, Immunofluorescence