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Thomas Gonatopoulos-Pournatzis Lab: Single-Cell Exon Deletion Profiling Reveals Splicing Events That Shape Gene Expression and Cell State Dynamics

Gonatopoulos-Pournatzis T
Unpublished

Plasmids from Article

ID Plasmid Purpose
237451pLCHKOv4Lentiviral backbone for expressing U6-driven hybrid guide (hg)RNAs for scCHyMErA-Seq experiments.
237452pLCHKOv5Lentiviral backbone for expressing doxycycline (Dox)-inducible, Pol II-driven hybrid guide (hg)RNAs.
237453pcDNA5-miniTurbo-NRF1-FLVector for generating Flp-In cell lines allowing dox-inducible mammalian expression of miniTurbo fused to full-length NRF1 isoform
237454pcDNA5-miniTurbo-NRF1-∆ex7Vector for generating Flp-In cell lines allowing dox-inducible mammalian expression of miniTurbo fused to NRF1 isoform lacking the alternative exon-7 (∆ex7)
237455pcDNA5-3xFLAG-NRF1-FL_WobbleMutVector for generating Flp-In cell lines allowing dox-inducible mammalian expression of 3xFlag-tagged full-length NRF1 isoform mutated to be resistant to NRF1 siRNA
237456pcDNA5-3xFLAG-NRF1-∆ex7_WobbleMutVector for generating Flp-In cell lines allowing dox-inducible mammalian expression of 3xFlag-tagged NRF1 isoform lacking the alternative exon-7 (∆ex7) mutated to be resistant to NRF1 siRNA
237457pCS2+ FlagVector for constitutive mammalian expression of gene of interest
237458pCS2+ Flag-SRSF3_WobbleMutVector for constitutive mammalian expression of Flag-tagged full-length SRSF3 mutated to be resistant to SRSF3 siRNA
237459pET01-NRF1-Ex7-minigeneVector for constitutive expression of NRF1 alternative exon-7 minigene
237460pX461-ABE8e-nSpCas9Transfection-based all-in-one base editor plasmid that expresses both adenine base editor (ABE8e-nSpCas9) and single guide RNAs (sgRNA) with BpiI cloning sites to clone sgRNA.
237461pX461-evoCDA1-nSpCas9Transfection-based all-in-one base editor plasmid that expresses both cytosine base editor (evoCDA1-nSpCas9) and single guide RNAs (sgRNA) with BpiI cloning sites to clone sgRNA.
237462pX462-ABE8e-nSaCas9Transfection-based all-in-one base editor plasmid that expresses both adenine base editor (ABE8e-nSaCas9) and single guide RNAs (sgRNA) with BpiI cloning sites to clone sgRNA.
237463pX461-ABE8e-nSpCas9-Intergenic sgRNAAll-in-one base editor expressing adenine base editor (ABE8e-nSpCas9) and gRNA control targeting Intergenic site
237464pX461-evoCDA1-nSpCas9-Intergenic sgRNAAll-in-one base editor expressing cytosine base editor (evoCDA1-nSpCas9) and sgRNA control targeting Intergenic site
237465pX462-ABE8e-nSaCas9-Intergenic sgRNAAll-in-one base editor expressing adenine base editor (ABE8e-nSaCas9) and sgRNA control targeting Intergenic site
237466pX461-ABE8e-nSpCas9-NRF1-ss-sgRNAAll-in-one adenine base editor (ABE8e-nSpCas9) with gRNA targeting NRF1-Ex7 splice site
237467pX461-evoCDA1-nSpCas9-NRF1-ss-sgRNAAll-in-one cytosine base editor (evoCDA1-nSpCas9) with sgRNA targeting NRF1-Ex7 splice site
237468pX462-ABE8e-nSaCas9-NRF1-ss-sgRNAAll-in-one adenine base editor (ABE8e-nSaCas9) with gRNA targeting NRF1-Ex7 splice site
237473scCHyMErA-Seq Exon Deletion Library 1Enables high-throughput identification of frame-preserving exons that influence transcriptional phenotypes, building on the CHyMErA system (Nature Biotechnology, 2020; PMID: 32249828).
237553Topo_SpCas9.tracrRNA-CS1_AsCas12a DRv10TOPO vector for the cloning of the SpCas9 tracrRNA-CS1 - AsCas12a Direct Repeat (DR) v10 fragment into the pLCHKO_v4 hgRNA vector for scCHyMErA-Seq experiments.

Antibodies from Article