Puromycin is an aminonucleoside antibiotic produced by the bacterium Streptomyces alboniger. It inhibits protein synthesis by disrupting peptide transfer on ribosomes causing premature chain termination during translation.

Puromycin, in conjunction with the CRISPR-CAS9 system, continues to be applied for genomic purposes. In this protocol, plasmid transfections utilizing these components will be described. PHASE 1. CRISPR Activation Plasmid Transient Transfection This protocol is recommended for a single well from a 6-well tissue culture plate. Adjust cell and reagent amounts proportionately for wells or dishes of different sizes. In...

Puromycin continues to be applied for genomic purposes. Continuing our puromycin series is a protocol on double nickase transfection. PHASE 1. Double Nickase Transfection This protocol is recommended for a single well from a 6-well tissue culture plate. Adjust cell and reagent amounts proportionately for wells or dishes of different sizes. In a 6-well tissue culture plate seed 1.5 x 105...

Puromycin continues to be applied for genomic purposes. In this protocol, puromycin is utilized in KO and HDR CRISPR plasmid transfections. PHASE 1. CRISPR/Cas9 KO Plasmid and HDR Plasmid Transfection This protocol is recommended for a single well from a 6-well tissue culture plate. Adjust cell and reagent amounts proportionately for wells or dishes of different sizes. In a 6-well tissue...

Puromycin continues to be applied for genomic purposes. In this protocol, puromycin is utilized in lentiviral transduction in shRNA particles. DAY 1 Plate target cells in a 12-well plate 24 hours prior to viral infection. Add 1 ml of complete optimal medium (with serum and antibiotics) and incubate cells overnight. The cells should be approximately 50% confluent on the day...

Puromycin's longstanding application as a gene selection antibiotic has a powerful new role in genome editing utilizing the CRISPR/Cas9 system. Here we discuss a protocol for shRNA Gene Silencing and CRISPR system transfection of which include puromycin. shRNA Plasmid DNA Mediated Inhibition of Gene Expression In a six well tissue culture plate, grow cells to a 50-70% confluency in antibiotic-free normal...

Puromycin N-acetyl transferase gene (pac), of which the gene product catalyzes antibiotic puromycin, an effective inhibitor of protein synthesis, has been widely used as a dominant selection marker in embryonic stem (ES) cell-mediated transgenesis.

Puromycin is an aminonucleosidic antibiotic. Puromycin is a protein synthesis inhibitor by inhibiting translation. Puromycin’s main function is used in cell biology as a selective antibiotic agent in cell culture systems.