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Genome-scale exon perturbation screens uncover exons critical for cell fitness

Xiao M-S, Damodaran AP, Kumari B, Dickson E, Xing K, On TA, Parab N, King HE, Perez AR, Guiblet WM, Duncan G, Che A, Chari R, Andresson T, Vidigal JA, Weatheritt RJ, Aregger M, Gonatopoulos-Pournatzis T
Mol Cell. 2024 June 24. doi: 10.1016/j.molcel.2024.05.024. (Link opens in a new window) Article

Plasmids from Article

ID Plasmid Purpose
209020pLenti-LbCas12a-8xNLSLentiviral vector expressing human codon-optimized LbCas12a with 8xNLS
209021pLenti-enCas12a-2xNLSLentiviral vector expressing human codon-optimized enCas12a with 2xNLS
209022pLenti-opCas12a-6xNLSLentiviral vector expressing human codon-optimized OpCas12a with 6xNLS
209023pLenti-UltraCas12a-6xNLSLentiviral vector expressing human codon-optimized UltraCas12a with 6xNLS
209024pLenti-opCas12a-Ultra-6xNLSLentiviral vector expressing human codon-optimized opCas12a-Ultra with 6xNLS
209025pLCHKOv3Lentiviral backbone for expressing U6 driven hybrid guide (hg)RNAs with BveI cloning sites and puromycin selection marker
209026pLCHKOv3-intergenic-AsLentiviral vector expressing U6 driven hybrid guide (hgRNA) targeting two intergenic sites in the human genome using SpCas9 and AsCas12a nucleases (CHyMErA system), respectively
209027pLCHKOv3-CD46 Ex3_1-LbLentiviral vector expressing U6 driven hybrid guide (hgRNA) targeting human CD46 Exon 3 for deletion using SpCas9 and LbCas12a nucleases (CHyMErA system)
209028pLCHKOv3-CD46 Ex3_2-LbLentiviral vector expressing U6 driven hybrid guide (hgRNA) targeting human CD46 Exon 3 for deletion using SpCas9 and LbCas12a nucleases (CHyMErA system)
209029pLCHKOv3-CD46 Ex3_3-LbLentiviral vector expressing U6 driven hybrid guide (hgRNA) targeting human CD46 Exon 3 for deletion using SpCas9 and LbCas12a nucleases (CHyMErA system)
209030pLCHKOv3-intergenic-LbLentiviral vector expressing U6 driven hybrid guide (hgRNA) targeting two intergenic sites in the human genome using SpCas9 and LbCas12a nucleases (CHyMErA system), respectively
209031pLCHKOv3-CD46 Ex3_1-AsLentiviral vector expressing U6 driven hybrid guide (hgRNA) targeting human CD46 Exon 3 for deletion using SpCas9 and AsCas12a nucleases (CHyMErA system)
209032pLCHKOv3-CD46 Ex3_2-AsLentiviral vector expressing U6 driven hybrid guide (hgRNA) targeting human CD46 Exon 3 for deletion using SpCas9 and AsCas12a nucleases (CHyMErA system)
209033pLCHKOv3-CD46 Ex3_3-AsLentiviral vector expressing U6 driven hybrid guide (hgRNA) targeting human CD46 Exon 3 for deletion using SpCas9 and AsCas12a nucleases (CHyMErA system)
209034pLCHKOv3-mClover3Lentiviral backbone for expressing U6 driven hybrid guide (hg)RNAs with BveI cloning sites, mClover3 and puromycin selection marker
209035pLCHKOv3-mCherryLentiviral backbone for expressing U6 driven hybrid guide (hg)RNAs with BveI cloning sites, mCherry and puromycin selection marker
209036pLCHKOv3-mClover-intergenic-AsLentiviral vector expressing mClover3 along with U6 driven hybrid guide (hgRNA) targeting two intergenic sites in the human genome using SpCas9 and AsCas12a nucleases (CHyMErA system), respectively
209037pLCHKOv3-mCherry-intergenic-AsLentiviral vector expressing mCherry along with U6 driven hybrid guide (hgRNA) targeting two intergenic sites in the human genome using SpCas9 and AsCas12a nucleases (CHyMErA system), respectively
209038pLenti-nSpCas9-NoCBELentiviral vector expressing doxycycline-inducible human codon-optimized SpCas9(D10A)-6xNLS (Cas9 nickase) linked to ssDNA binding domain from RAD51 and uracil DNA glycosylase inhibitor (UGI)
209039pLenti-nSpCas9-APOBEC1Lentiviral vector expressing doxycycline-inducible human codon-optimized cytosine base editor where APOBEC1 is fused to the N-terminus of SpCas9(D10A)-sDNA Rad51-2xUGI-6xNLS
209040pLenti-nSpCas9-APOBEC3ALentiviral vector expressing doxycycline-inducible human codon-optimized cytosine base editor where APOBEC3A is fused to the N-terminus of SpCas9(D10A)-sDNA Rad51-2xUGI-6xNLS
209041pLenti-nSpCas9-BEACON2Lentiviral vector expressing doxycycline-inducible human codon-optimized cytosine base editor where BEACON2 is fused to the N-terminus of SpCas9(D10A)-sDNA Rad51-2xUGI-6xNLS
209042pLenti-nSpCas9-evoCDA1Lentiviral vector expressing doxycycline-inducible human codon-optimized cytosine base editor where evoCDA1 is fused to the N-terminus of SpCas9(D10A)-sDNA Rad51-2xUGI-6xNLS
209043pLenti-nSpCas9-NoABELentiviral vector expressing doxycycline-inducible human codon-optimized SpCas9(D10A)-6xNLS (Cas9 nickase)
209044pLenti-nSpCas9-TadA8eLentiviral vector expressing doxycycline-inducible human codon-optimized adenine base editor where TadA8e (ABE8e) is fused to the N-terminus of SpCas9(D10A)-6xNLS
209045pLenti-nSpCas9-TadA8.20mLentiviral vector expressing doxycycline-inducible human codon-optimized adenine base editor where TadA8.20m is fused to the N-terminus of SpCas9(D10A)-6xNLS
209046pLCKOv3Lentiviral backbone for expressing U6 driven SpCas9 sgRNAs with BveI cloning sites, SpCas9 tracrRNAv1 (Dang et al., 2015; PMID: 26671237) and puromycin selection marker
209047pDONR223-TAF5-FLGateway-compatible entry vector, with insert of the full length TAF5 gene
209048pDONR223-TAF5-∆ex8Gateway-compatible entry vector, with insert of the TAF5 isoform lacking the alternative exon-8 (∆ex8)
209049pcDNA5-3xFLAG-TAF5-FLVector for generating Flp-In cell lines allowing dox-inducible mammalian expression of 3xFlag-tagged full length TAF5 isoform
209050pcDNA5-3xFLAG-TAF5-∆ex8Vector for generating Flp-In cell lines allowing dox-inducible mammalian expression of 3xFlag-tagged TAF5 isoform lacking the alternative exon-8 (∆ex8)
209051pcDNA5-miniTurbo-TAF5-FLVector for generating Flp-In cell lines allowing dox-inducible mammalian expression of miniTurbo fused to full length TAF5 isoform
209052pcDNA5-miniTurbo-TAF5-∆ex8Vector for generating Flp-In cell lines allowing dox-inducible mammalian expression of miniTurbo fused to TAF5 isoform lacking the alternative exon-8 (∆ex8)
209053pDONR223-TAF5-FL_WobbleMutGateway-compatible entry vector, with insert of the full length TAF5 gene mutated to be resistant to TAF5 siRNA
209054pDONR223-TAF5-∆ex8_WobbleMutGateway-compatible entry vector, with insert of the TAF5 isoform lacking the alternative exon-8 (∆ex8) mutated to be resistant to TAF5 siRNA
209055pcDNA5-3xFLAG-TAF5-FL_WobbleMutVector for generating Flp-In cell lines allowing dox-inducible mammalian expression of 3xFlag-tagged full length TAF5 isoform mutated to be resistant to TAF5 siRNA
209056pcDNA5-3xFLAG-TAF5-∆ex8_WobbleMutVector for generating Flp-In cell lines allowing dox-inducible mammalian expression of 3xFlag-tagged TAF5 isoform lacking the alternative exon-8 (∆ex8) mutated to be resistant to TAF5 siRNA
209057pDONR223-EGFPGateway-compatible entry vector, with insert of the EGFP gene
209058pcDNA5-3xFLAG-EGFPVector for generating Flp-In cell lines allowing dox-inducible mammalian expression of 3xFlag-tagged EGFP
209059pcDNA5-miniTurbo-EGFPVector for generating Flp-In cell lines allowing dox-inducible mammalian expression of miniTurbo fused to EGFP
209734CHyMErA LbCas12a Nuclease Optimization hgRNA LibraryCHyMErA Cas12a Nuclease Optimization hgRNA Libraries allows assessment and comparison of the efficacy of different LbCas12a nucleases in the context of the Cas9-Cas12a CHyMErA system.
209735CHyMErA AsCas12a Nuclease Optimization hgRNA LibraryCHyMErA Cas12a Nuclease Optimization hgRNA Libraries allows assessment and comparison of the efficacy of different AsCas12a nucleases in the context of the Cas9-Cas12a CHyMErA system.
209736CHyMErA Large-Scale Exon Deletion hgRNA LibraryCHyMErA Exon Deletion hgRNA Library allows identification of frame-preserving exons that affect cell fitness in human cells using the CHyMErA screening platform.
218814pLCHKOv3-mClover-intergenic_2-AsLentiviral vector expressing mClover3 along with U6 driven hybrid guide (hgRNA) targeting two intergenic sites in the human genome using SpCas9 and AsCas12a nucleases (CHyMErA system), respectively
218815pLCHKOv3-mClover-intergenic_3-AsLentiviral vector expressing mClover3 along with U6 driven hybrid guide (hgRNA) targeting two intergenic sites in the human genome using SpCas9 and AsCas12a nucleases (CHyMErA system), respectively
218816pLCHKOv3-mClover-intergenic_4-AsLentiviral vector expressing mClover3 along with U6 driven hybrid guide (hgRNA) targeting two intergenic sites in the human genome using SpCas9 and AsCas12a nucleases (CHyMErA system), respectively
218972PET01-TAF5-ex8-Minigene-SRSF1 motif WTVector for constitutive expression of TAF5 alternative exon-8 mini gene with wild type SRSF1 motif (TCAGAGGA)
218973pET01-TAF5-ex8-Minigene-SRSF1 motif MUTVector for constitutive expression of TAF5 alternative exon-8 mini gene with mutated SRSF1 motif (AGTCTCCT)
218974pcDNA5-3xFLAG-SRSF1 siRNA-resitantVector for expressing siRNA-resistant SRSF1 cDNA (siRNA sequence: CGUGGAGUUUGUACGGAAA)

Antibodies from Article