pcas9 cat (Addgene inc)

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Addgene inc pcas9 cat
Pcas9 Cat, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 60 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/pcas9+cat/pSAG1%3A%3ACAS9-U6%3A%3AsgUPRT+(Plasmid+%2354467)/bio_rxiv__64898__2026__01__22__701003-272-14-15
Average 93 stars, based on 60 article reviews

pcas9 cat - by Bioz Stars, 2026-09

93/100 stars

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Transfection:

Article Title: Differentiation of Toxoplasma into latent forms is linked to central carbon metabolism and requires a GID/CTLH-type E3 ubiquitin ligase
Article Snippet: Parasites were resuspended in the DNA mixture and electroporated using a Lonza nucleofector, using program F1-115. .. To create the RH: cas9 strain, RH:Δ hxgprt parasites were transfected simultaneously with pCas9-CAT (Addgene # 80323) and pU6-Decoy (Addgene #80324) plasmids together, as done previously . .. To create Pru: cas9 , Pru:Δ hxgprt parasites were transfected with linearized pBM019 (Addgene #128179) plasmid that encodes Cas9 .

FLAG-tag:

Article Title: Helical Repeat Protein mRSiC is Required for rRNA Fragment Accumulation in T. gondii Mitochondria
Article Snippet: For C-terminal tagging of mRSiC (TGME49 237530), a sgRNA targeting the 3’ end of mRSiC was introduced into pSAG1::Cas9-U6::sgUPRT (Addgene plasmid # 54467; ( Shen et al ., 2014 ) by using the Q5-site directed mutagenesis kit (NEB), following the manufacturer’s instructions. .. A short glycine linker followed by a 3x FLAG tag sequence was integrated into pCas9-CAT (Addgene plasmid # 54467; ( Sidik et al ., 2016 ) and the resulting linker/3xFLAG/CAT cassette amplified using Q5 polymerase (NEB) and primers containing 50 bp overhangs homologous to the either upstream or downstream regions of the mRSiC stop codon. ..

Sequencing:

Plasmid Preparation:

Article Title: CRISPR/Cas9-based genome-wide screening of Toxoplasma gondii
Article Snippet: .. We have since been able to generate similar strains with a single plasmid containing the Cas9, a selectable marker, and the decoy sgRNA. pCas9-CAT and pU6-Decoy are available from Addgene, and RH/Cas9 will soon be available from the ATCC. .. Investigators wishing to generate their own Cas9-expressing strains may use the plasmids available from Addgene, and should take care to characterize the efficiency of gene disruption in a clonal line of parasites that homogenously expresses Cas9.

Amplification:

Marker:

CRISPR:

Article Title: CRISPR/Cas9-based genome-wide screening of Toxoplasma gondii
Article Snippet: Petri dishes (Falcon cat. no. 351029) Culture tubes (Corning cat. no. 352051) 24-well tissue culture dishes (Costar cat. no. 3524) Parafilm (VWR cat. no. 291–1214) T12.5 tissue culture flasks (Falcon cat. no. 353018) 15 cm tissue culture dishes (Falcon cat. no. 353025) Cell scrapers (Costar cat. no. 3010) PCR tubes (BioRad cat. no. TF102010) Cryotubes (Thermo Scientific cat. no. 5000–0012) 271⁄2 -gauge needles (BD cat. no. 305109) CAUTION Exercise extreme care when handling live T. gondii with sharps. .. Blunt needles (SAI Infusion cat. no. B27–50) can be used as an alternative. pU6-SAG1 (Addgene cat. no. 80322) Lourido Toxoplasma CRISPR Library V1 (Addgene cat. no. 80636) pU6-DHFR (Addgene cat. no. 80329) pCas9-CAT (Addgene cat. no. 80323) pU6-Decoy (Addgene cat. no. 80324) Ampicillin (Sigma-Aldrich-Aldrich cat. no. A0166–25G) Glycerol (EMD cat. no. GX0185–6) Methanol-Free Formaldehyde (Thermo Scientific cat. no. 28908) Prolong Gold (Invitrogen cat. no. P1044) Molecular biology-grade water (e.g., from an EMD Millipore Milii-Q Integral System, Fisher Scientific cat. no. ZRXQ003US) Gentamicin (Gibco cat. no. 15710–064) EGTA (Sigma-Aldrich cat. no. E4378–500G) HEPES (Sigma-Aldrich cat. no. 3375–1KG) KH 2 PO 4 (Sigma-Aldrich cat. no. P5655) K 2 HPO 4 (Sigma-Aldrich cat. no. 1551128) KCl (Fisher Scientific cat. no. SP138–500) MgCl 2 (Mallinckrodt cat. no. 5958) Methanol for HPLC gradient analysis (Millipore, cat. No. 67–56-1) EDTA (Amresco cat. no. 0322–500G) Tris base (Sigma-Aldrich cat. no. TRIS-RO) Glacial acetic acid (Spectrum cat. no. A1010) CAUTION Glacial acetic acid is corrosive; avoid contact with eyes, skin, and clothing; wear protective gloves while handling. .. Na 2 HPO 4 (Sigma-Aldrich cat. no. 255793) NaCl (Sigma Aldrich cat. no. S7653) Glutathione (Sigma-Aldrich cat. no. G6013–5G) Adenosine Triphosphate (ATP, Sigma-Aldrich cat. no. A6419–1G) CaCl 2 (Sigma-Aldrich cat. no. C5080–1KG) Chloramphenicol (Sigma-Aldrich cat. no. C3175–100MG) Pyrimethamine (Sigma-Aldrich cat. no. 46706–250MG) NaOH (Sigma-Aldrich cat. no. S2770) CAUTION Sodium hydroxide is a strong base that can cause severe skin burns and eye injuries; avoid contact with eyes, skin and clothing; use in a fume hood and wear protective eyewear and gloves during handling.

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