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An engineered transcriptional reporter of protein localization identifies regulators of mitochondrial and ER membrane protein trafficking in high-throughput CRISPRi screens.

Coukos RW, Yao D, Sanchez ML, Strand ET, Olive ME, Udeshi ND, Weissman JS, Carr SA, Bassik MC, Ting AY
Elife. 2021 Aug 20;10. pii: 69142. doi: 10.7554/eLife.69142. (Link opens in a new window) PubMed (Link opens in a new window) Article

Plasmids from Article

ID Plasmid Purpose
170994UAS:mCherryUAS:GAL4 repeat-containing promoter drives simple mCherry reporter gene. Blasticidin selectable.
170995AKAP1-hLOV-TEVcs-GAL4bd-VP64-V5HiLITR transcription factor with AKAP1 N-terminal tmd targeting sequence (mitochondria)
170996TMED3-hLOV-TEVcs-GAL4bd-VP64-V5HiLITR transcription factor with full-length TMED3 targeting sequence (ER/Golgi)
170997PMP34-hLOV-TEVcs-GAL4bd-VP64-V5HiLITR transcription factor with full-length PMP34 targeting sequence (Peroxisome)
170998EGFP-uTEV1-MAVSHiLITR protease with MAVS c-terminal tmd targeting information (Mitochondria)
170999EGFP-uTEV1-MAVS-mTAHiLITR protease with MAVS [R537A] C-terminal tmd targeting information (Mitochondria and ER insertion)
171000EGFP-uTEV1-MAVS-mTA2HiLITR protease with MAVS [R537A, R538A] C-terminal tmd targeting information (Mitochondria and ER insertion)
171001EGFP-uTEV1-MAVS-mTA3HiLITR protease with MAVS [Y536delinsFIV] C-terminal tmd targeting information (Mitochondria and ER insertion)
171002EGFP-uTEV1-MAVS-mTA4HiLITR protease with MAVS [Y536delinsFIVLI] C-terminal tmd targeting information (Mitochondria and ER insertion)
171003EGFP-uTEV1-MAVS-mTA5HiLITR protease with MAVS [Y536delinsFIVLI, R537A] C-terminal tmd targeting information (Mitochondria and ER insertion)
171004EGFP-uTEV1-MAVS-mTA6HiLITR protease with MAVS [Y536delinsFIVLI, R537A, R538A] C-terminal tmd targeting information (Mitochondria)
171005AKAP1-EGFP-uTEV1HiLITR protease with AKAP1 N-terminal tmd targeting information (Mitochondria)
171006EGFP-uTEV1-NESHiLITR protease with C-terminal NNES (Cytoplasmic)
171007EGFP-uTEV1-CB5HiLITR protease with CB5 C-terminal tmd targeting information (ER)
171008EGFP-uTEV1-ACBD5HiLITR protease with ACBD5 C-terminal targeting information (peroxisome)
171009EGFP-uTEV1-SQStmdHiLITR protease with SQS C-terminal targeting information (ER)
171010EGFP-uTEV1-SQSflHiLITR protease with full-length SQS targeting information (ER)
171011pMCB619CRISPR sgRNA vector (puro-BFP with BstXI/BlpI digest sites).
171012pLX208-SAE1Ectopic expression of SAE1
171013pLX208-nfBFPEctopic expression of nonfluorescent BFP

Antibodies from Article