Plenti abera p2a puro

Manufactured by Addgene

The PLenti-ABERA-P2A-Puro is a plasmid vector that can be used for lentiviral transduction and expression of genes of interest in mammalian cells. The vector contains the ABERA gene, a P2A self-cleaving peptide, and a puromycin resistance cassette for selection of transduced cells.

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Lab products found in correlation

Nucleobond xtra midi ef, by Macherey-Nagel (2 mentions) Plenti fnls p2a puro, by Addgene (2 mentions) Exonuclease 1, by New England Biolabs (1 mentions) Pspax2, by Addgene (1 mentions) Pmd2 g, by Addgene (1 mentions)

2 protocols using plenti abera p2a puro

Construction of Lentiviral Vectors for CRISPR/Cas9 Editing

Cited 2 times

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The plasmids used in this study, pLenti-FNLS-P2A-Puro (Addgene#110841-CBE) and pLenti-ABERA-P2A-Puro (Addgene#112675-ABE), were a gift from Lukas Dow (Zafra et al., 2018 (link)), and pMD2.G and psPAX2 (second-generation lentiviral packaging construct, Addgene #12259, 11260) were a gift from Didier Trono. The pLenti-ABE8e-puro vector was constructed by amplifying ABE8e from the TadA-8e V106W plasmid (a plasmid gifted from David Liu, Addgene#138495) (Richter et al., 2020 (link)) using the primers mentioned in Supplementary file 2. The amplified PCR product was then cloned into pLenti-ABERA-P2A-Puro backbone after digestion with BamH1 and Nhe1 by HIFI assembly (NEB). The gRNA sequence from lentiCRISPR V2 vector (a construct gifted from Feng Zhang, Addgene#52961) was removed by digestion with EcoR1 and Kpn1 enzyme (NEB). The digested plasmid was then re-ligated with NEB Ligase after a exonuclease treatment (NEB Exonuclease 1) to generate a lentiCRISPR V2.1 vector (Sanjana et al., 2014 (link)). The plasmids were isolated using NucleoBond Xtra Midi EF (Macherey-Nagel) according to the manufacturer’s instruction.

Ravi N.S., Wienert B., Wyman S.K., Bell H.W., George A., Mahalingam G., Vu J.T., Prasad K., Bandlamudi B.P., Devaraju N., Rajendiran V., Syedbasha N., Pai A.A., Nakamura Y., Kurita R., Narayanasamy M., Balasubramanian P., Thangavel S., Marepally S., Velayudhan S.R., Srivastava A., DeWitt M.A., Crossley M., Corn J.E, & Mohankumar K.M. (2022). Identification of novel HPFH-like mutations by CRISPR base editing that elevate the expression of fetal hemoglobin. eLife, 11, e65421.

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Lentiviral Plasmid Isolation Protocol

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The plasmid in this study, pLenti-FNLS-P2A-Puro (Addgene#110841-CBE) and pLenti-ABERA-P2A-Puro (Addgene#112675-ABE ) was a gift from Lukas Dow (42), pMD2.G and psPAX2 (Addgene #12259, 11260-2nd generation lentiviral packaging construct )was a gift from Didier Trono. The plasmids were isolated using NucleoBond Xtra Midi EF (Macherey-Nagel) according to the manufacturer's instruction.

Ravi N.S., Wienert B., Wyman S.K., Vu J.T., Pai A.A., Balasubramanian P., Nakamura Y., Kurita R., Marepally S., Thangavel S., Velayudhan S.R., Srivastava A., DeWitt M.A., Corn J.E., & Murugesan M. (2020). Identification of novel HPFH-like mutations by CRISPR base editing that elevates the expression of fetal hemoglobin.

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