U2OS cells were transfected with a pool of 4 siRNAs (SMARTpool) targeting p21 (CDKN1a), sequences: GAUGGAACUUCGACUUUGU, GCGAUGGAACUUCGACUUU, CGAUGGAACUUCGACUUUG and CGACUGUGAUGCGCUAACG (Sigma). Alternatively, an independent p21 siRNA was used when indicated (p21siRNA #2); Mission siRNA CDK1NA (p21), sequence: CUAAGAGUGCUGGGCAUUU (Sigma). Cells were subjected to treatment 48 hours post transfection. The HCT116 wild type (wt) and p21 negative (p21-/-) cell lines were purchased from Horizon Discovery (HD R02-035), and p21 loss was verified by measuring p21 induction after radiation. RPE cells deficient for p21 were made by the CRISPR/cas9 method: To knock-out the CDKN1A gene, hTERT RPE-1 cells were transfected with a combination (1:1) of p21 CRISPR/Cas9 KO Plasmid (Santa Cruz, sc-400,013), and p21 HDR Plasmid and stable clones were selected by puromycin (10 μg/ml). Integration of the HDR cassette to the CDKN1A locus was confirmed by sequencing, and loss of p21 expression by immunoblotting.
P21 crispr cas9 ko plasmid
Manufactured by Santa Cruz Biotechnology
The P21 CRISPR/Cas9 KO Plasmid is a laboratory tool that enables targeted gene knockout using the CRISPR/Cas9 system. The plasmid contains the Cas9 endonuclease and a guide RNA targeting the p21 gene. This product can be used to disrupt the p21 gene in cell lines or other biological systems for research purposes.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using p21 crispr cas9 ko plasmid
1
p21 Knockdown in Cell Lines
2
Generation of p21 Knockout and Overexpressing A549 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A549 cells were transfected with a p21 CRISPR/Cas9 KO plasmid (#sc‐400013) and p21 HDR plasmid (#sc‐400013‐HDR) (Santa Cruz Biotechnology) using Lipofectamine 3000 (Invitrogen). Thereafter, p21‐knockout A549 (A549‐KOp21) cell colonies were selected using puromycin (Sigma‐Aldrich). To establish p21‐expressing cells from A549‐KOp21 cells, pEB Multi‐Neo vector (Fujifilm Wako Pure Chemical), in which a human p21 gene was inserted, was transfected into A549‐KOp21 cells using Lipofectamine 3000 (Invitrogen), followed by culture with G418 (Nacalai Tesque). To establish p21‐overexpresing A549 cells, parental A549 cells were similarly transfected with the p21‐encoding pEB Multi‐Neo vector.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!