PTRF CRISPR/Cas9 KO Plasmid

Protocol tips

Upstream tips	Protocol tips	Downstream tips
If transfecting more than one plasmid (i.e., CRISPR/Cas9 KO Plasmid with HDR Plasmid), mix Plasmid DNA at equivalent ratios.	Do not add antibiotics to the Plasmid Transfection Medium

Upstream tips
If transfecting more than one plasmid (i.e., CRISPR/Cas9 KO Plasmid with HDR Plasmid), mix Plasmid DNA at equivalent ratios.
Protocol tips
Do not add antibiotics to the Plasmid Transfection Medium

Publication protocol

Generation of PTRF null 3T3-L1 stable cell lines by CRISPR/Cas9 genome editing
The PTRF null 3T3-L1 stable cell lines were generated using lentiviral plasmid based CRISPR/Cas9 genome editing, in which U6 promoter driven guide RNA, CMV promoter driven cas9 and puromycin selection marker were supplied by one single-vector (all-in-one). Four vectors, including three targets (K1: 5’-TCACGCTCCATATCGTTGAG-3’; K2: 5’-GTCAACGTGAAGACCGTGCG-3’; K3: 5’-GGTCAGCTGGATCTGGTCAA-3’) and one non-targeting control were custom designed and made by Transomic Technologies (Huntsville, AL). Lentivirus were packing used third generation packing system. The stable cell lines were obtained after lentivirus transduction and puromycin selection. These cells were cultured and differentiated as described above.



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Manufacturer protocol

Download the product protocol from Santa Cruz Biotechnology for PTRF CRISPR/Cas9 KO Plasmid below.

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