New plasmids from us deposited to AddGene

We have deposited two plasmids generated by drs. Sudeepta Kumar Panda and Sanjay Boddul, used in our recent PNAS paper (Panda et al, PNAS, 2020), to Addgene.

The first plasmid is a development of the LentiGuide-Puro from the Zhang lab where Sudeepta inserted a P2A-EGFP cassette enabling both Puro selection and EGFP based selection. We typically use the EGFP signal to evaluate the MOI and monitor the efficiency of puro based selection:


The second plasmid was developed from the LentiGuide-Puro-P2A-EGFP by Sanjay to also include an RFP reporter in the stuffer region driven by a bacterial promoter. This enables you to rapidly evaluate if bacteria expanded on agar/amp plates have taken up constructs where the stuffer is intact or not. This is very helpful to evaluate how well the cloning of a CRISPR/gRNA library, replacing the stuffer, into the plasmid has worked. A big thanks to the molecular biology wizard Erik Holmgren for help with the mRFP1 sequence.

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