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Bsmbi cut lenticrispr v2 plasmid

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The BsmBI cut lentiCRISPR v2 plasmid is a tool for gene editing. It contains the necessary elements for CRISPR-Cas9 mediated genome engineering using lentiviral transduction.

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LentiCRISPR v2 (605 citations) LentiCRISPR v2 plasmid (170 citations) LentiCRISPR plasmid (21 citations) LentiCRISPR (11 citations)

2 protocols using bsmbi cut lenticrispr v2 plasmid

1

CRISPR-mediated Knockout of Mouse SLC38A2

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sgRNAs targeting the first exon of mouse SLC38A2 were identified using http://crispr.mit.edu/online tool and subcloned into BsmBI cut lentiCRISPR v2 plasmid (Addgene, 52961) (Sanjana et al., 2014 (link)) using a set of primers to anneal (CAC CGT TCA ACA TCT CCC CGG ATG; AAA CCA TCC GGG GAG ATG TTG AAC). Insert position was verified by sequencing. Propagation of lentiviral particles expressing lentiCRISPR sgSLC38A2 was performed in HEK293T cells. S51A MEFs were infected with the lentivirus and 24 h later, cells were selected with puromycin (2.5 μg/mL) for 3 days. The deletion was verified by western blotting.
Krokowski D., Jobava R., Szkop K.J., Chen C.W., Fu X., Venus S., Guan B.J., Wu J., Gao Z., Banaszuk W., Tchorzewski M., Mu T., Ropelewski P., Merrick W.C., Mao Y., Sevval A.I., Miranda H., Qian S.B., Manifava M., Ktistakis N.T., Vourekas A., Jankowsky E., Topisirovic I., Larsson O, & Hatzoglou M. (2022). Stress-induced perturbations in intracellular amino acids reprogram mRNA translation in osmoadaptation independently of the ISR. Cell reports, 40(3), 111092.
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2

CRISPR-mediated Knockout of Mouse SLC38A2

Check if the same lab product or an alternative is used in the 5 most similar protocols
sgRNAs targeting the first exon of mouse SLC38A2 were identified using http://crispr.mit.edu/online tool and subcloned into BsmBI cut lentiCRISPR v2 plasmid (Addgene, 52961) (Sanjana et al., 2014 (link)) using a set of primers to anneal (CAC CGT TCA ACA TCT CCC CGG ATG; AAA CCA TCC GGG GAG ATG TTG AAC). Insert position was verified by sequencing. Propagation of lentiviral particles expressing lentiCRISPR sgSLC38A2 was performed in HEK293T cells. S51A MEFs were infected with the lentivirus and 24 h later, cells were selected with puromycin (2.5 μg/mL) for 3 days. The deletion was verified by western blotting.
Krokowski D., Jobava R., Szkop K.J., Chen C.W., Fu X., Venus S., Guan B.J., Wu J., Gao Z., Banaszuk W., Tchorzewski M., Mu T., Ropelewski P., Merrick W.C., Mao Y., Sevval A.I., Miranda H., Qian S.B., Manifava M., Ktistakis N.T., Vourekas A., Jankowsky E., Topisirovic I., Larsson O, & Hatzoglou M. (2022). Stress-induced perturbations in intracellular amino acids reprogram mRNA translation in osmoadaptation independently of the ISR. Cell reports, 40(3), 111092.
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