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Minimal genetically encoded tags for fluorescent protein labeling in living neurons.
Arsic A, Hagemann C, Stajkovic N, Schubert T, Nikic-Spiegel I
Nat Commun. 2022 Jan 14;13(1):314. doi: 10.1038/s41467-022-27956-y. PubMed Article

Plasmids from Article

ID Plasmid Purpose
182287pcDNA3.1(+)_U6 tRNAPyl_CMV NESPylRS(AF) Encodes codon-optimized AF mutant of M. mazei-derived pyrrolysine (Pyl) tRNA synthetase fused to a nuclear export signal (NESPylRSAF) & tRNACUAPyl used for amber codon suppression in mammalian cells
182652pcDNA3.1(+)_4x(U6 tRNA M15)_CMV NESPylRS(AF)Encodes codon-optimized AF mutant of M. mazei pyrrolysine (Pyl) tRNA synthetase fused to a nuclear export signal (NESPylRSAF) & 4x M15 tRNACUA used for amber codon suppression in mammalian cells
182653pcDNA3.1(+)_U6 tRNAPyl_CMV NESPylRS(AF)_IRES_eRF1(E55D)-HAEncodes codon-optimized AF mutant of M. mazei pyrrolysine (Pyl) tRNA synthetase fused to a nuclear export signal (NESPylRSAF),tRNACUAPyl & eRF1E55D used for amber codon suppression in mammalian cells
182654pCMV_NFL(WT)-FLAGExpresses wild-type mouse neurofilament light chain with a C-terminal FLAG tag (DYKDDDDK) in mammalian cells.
182655pCMV_NFL(K211TAG)-FLAGExpresses mouse neurofilament light chain with a TAG codon at the position K211 and a C-terminal FLAG tag (DYKDDDDK) in mammalian cells.
182656pCMV_NFL(K363TAG)-FLAGExpresses mouse neurofilament light chain with a TAG codon at the position K363 & C-terminal FLAG tag (DYKDDDDK) in mammalian cells. Can be used for amber codon suppression & click labeling of NFL.
182657pCMV_NFL(R438TAG)-FLAGExpresses mouse neurofilament light chain with a TAG codon at the position R438 and a C-terminal FLAG tag (DYKDDDDK) in mammalian cells.
182658pCMV_NFL(K468TAG)-FLAGExpresses mouse neurofilament light chain with a TAG codon at the position K468 and a C-terminal FLAG tag (DYKDDDDK) in mammalian cells.
182659pCMV_FLAG-NFL(WT)Expresses wild-type mouse neurofilament light chain with an N-terminal FLAG tag (DYKDDDDK) in mammalian cells.
182660pCMV_FLAG-NFL(K211TAG)Expresses mouse neurofilament light chain with a TAG codon at the position K211 and an N-terminal FLAG tag (DYKDDDDK) in mammalian cells.
182661pCMV_FLAG-NFL(K363TAG)Expresses mouse neurofilament light chain with a TAG codon at the position K363 & an N-terminal FLAG tag (DYKDDDDK) in mammalian cells. Can be used for genetic code expansion & click labeling of NFL.
182662pCMV_FLAG-NFL(R438TAG)Expresses mouse neurofilament light chain with a TAG codon at the position R438 and an N-terminal FLAG tag (DYKDDDDK) in mammalian cells.
182663pCMV_FLAG-NFL(K468TAG)Expresses mouse neurofilament light chain with a TAG codon at the position K468 and an N-terminal FLAG tag (DYKDDDDK) in mammalian cells.
182678pORANGE_Tubb3 GFP(Y39TAG) KIExpression of spCas9, gRNA targeting the end of Tubb3 gene and donor GFP(Y39TAG). Can be used for amber codon suppression and click chemistry labeling of endogenous beta-3 tubulin in mammalian cells.
182679pORANGE_NFL linker-3xFLAG KIExpression of spCas9, gRNA targeting the end of mouse Nefl gene and donor linker-3xFLAG. Can be used for C-terminal tagging of endogenous NFL with a linker-3xFLAG tag.
182680pORANGE_NFL linker(A6TAG)-3xFLAG KIExpression of spCas9, gRNA targeting the end of mouse Nefl gene and donor linker(A6TAG)-3xFLAG. Can be used for amber codon suppression and click chemistry labeling of endogenous NFL.
182681TKIT NFL gRNA1 gRNA2Expression of spCas9, gRNA targeting intron 3 and gRNA targeting 3'UTR of the mouse Nefl gene. Can be used for the tagging of endogenous NFL via Targeted Knock-In with Two guides (TKIT) approach.
182682pcDNA3.1/Zeo(+)_TKIT NFL intron3/exon4 linker(WT)-3xFLAG donorContains a donor sequence with a part of Nefl intron 3, whole Nefl exon 4 and linker-3xFLAG. Used as a donor for endogenous NFL tagging via Targeted Knock-In with Two guides (TKIT) approach.
182685pcDNA3.1/Zeo(+)_mCherry_TKIT NFL intron3/exon4 linker(WT)-3xFLAG donorEncodes mCherry and contains a donor sequence with a part of Nefl intron 3, whole Nefl exon 4 and linker-3xFLAG. Can be used as a donor plasmid for the tagging of endogenous NFL via TKIT approach.
182686pcDNA3.1/Zeo(+)_TKIT NFL intron3/exon4 linker(TAG)-3xFLAG donorContains a donor sequence with a part of Nefl intron 3, whole Nefl exon 4 and linkerA6TAG-3xFLAG. Can be used as a donor for the tagging of endogenous NFL via TKIT approach and click labeling of NFL.

Antibodies from Article