Pmscarlet i c1

Manufactured by Addgene

PmScarlet-i_C1 is a red fluorescent protein variant derived from mScarlet. It exhibits enhanced brightness and photostability compared to other red fluorescent proteins. The core function of PmScarlet-i_C1 is to serve as a fluorescent marker for various biological applications.

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Lab products found in correlation

Pce mp53dd, by Addgene (1 mentions) Pspax2, by Addgene (1 mentions) Pmd2 g, by Addgene (1 mentions) Hcas9, by Addgene (1 mentions) Pspcas9 bb 2a puro px459 v2, by Addgene (1 mentions) Aavs1 puro pgk1 3xflag twin strep, by Addgene (1 mentions) Pfuultra 2 fusion hotstart dna polymerase, by Agilent Technologies (1 mentions) Memerald sec61β, by Addgene (1 mentions) Gblock, by Integrated DNA Technologies (1 mentions)

Close spelling variants of this product

PmScarlet_C1

4 protocols using pmscarlet i c1

Plasmid Constructs for Cellular Imaging

Cited 4 times

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pMSCV-Puro-EYFP-vinculin, pMSCV-Puro-EYFP-mCasWT, pMSCV-Puro-EYFP-mCas15F, and pBabe-Puro-mCherry-mVCL were described previously (Teckchandani and Cooper, 2016 (link)). EGFP-Rac1^WT and EGFP-Rac1^Q61L were provided by K. Wennerberg, University of North Carolina, Chapel Hill, NC (Arthur et al., 2004 (link)). The following vectors were gifts from the indicated investigators: pmScarlet-i_C1 (Dorus Gadella, Addgene plasmid # 85044), pORANGE cloning template vector (Harold MacGillavry, Addgene # 131471), pCE-mp53DD (Shinya Yamanaka, Addgene # 41856), pLenti-Rac1-2G (Olivier Pertz, Addgene # 66111), pcDNA3.1-mGreenLantern (Gregory Petsko, Addgene # 161912) (Campbell et al., 2020 (link)), pMD2.G and psPAX2 (Didier Trono, Addgene #12259 and 12260). pLenti Ecto-pHluorin β1 integrin with 4-residue linkers was kindly provided by David A. Calderwood (Yale University School of Medicine, USA) (Huet-Calderwood et al., 2017 (link)).

Kumar S., Stainer A., Dubrulle J., Simpkins C, & Cooper J.A. (2023). Cas phosphorylation regulates focal adhesion assembly. eLife, 12, e90234.

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LDAF1-FLAG-Seipin(1-310) Complex Expression

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The following plasmids were kind gifts: ERoxBFP (Addgene plasmid #68126) from Erik Snapp, mEmerald-Sec61β (Addgene plasmid #54249) from Michael Davidson, hCas9 (Addgene plasmid #41815) and gRNA-AAVS1-T2 (Addgene plasmid #41818) from George Church, AAVS1_Puro_PGK1_3xFLAG_Twin_Strep (Addgene plasmid #68375) from Yannick Doyon, pSpCas9(BB)-2A-Puro (PX459) V2.0 (Addgene plasmid #62988) from Feng Zhang, and pmScarlet-i_C1 (Addgene plasmids #85044) from Dorus Gadella. pEGFP-N1 and pEGFP-C1 plasmids were purchased from Clontech Laboratories, pSMART-HC-Amp plasmid was purchased from Lucigen. pCAG-LNK vector was modified from pCAGEN (Addgene plasmid #11160) as described (Scheich et al., 2007).
For plasmid construction, all PCRs were performed using PfuUltra II Fusion HotStart DNA Polymerase (#600672, Agilent Technologies) and restriction enzymes were from New England Biolabs. The synthetic DNAs (gBlock, Integrated DNA Technologies) that were used in this study and cloning strategies of the other plasmids (including primer information) were summarized in Table S3 and S4, respectively.
For expression of the LDAF1-FLAG-seipin(1-310) complex, pCAG-LDAF1-FLAG-Seipin(1-310) plasmid was generated by pCAG-LNK-LDAF1-FLAG and pCAG-LNK-Seipin(1-310) using approach described in Scheich et al.(Scheich et al., 2007). The detailed cloning strategies were summarized in Table S3 and S4.

Chung J., Wu X., Lambert T.J., Lai Z.W., Walther T.C, & Farese RV J.r. (2019). Lipid droplet assembly factor-1 and seipin form a lipid droplet assembly complex. Developmental cell, 51(5), 551-563.e7.

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Cloning and Characterization of mScarlet-I Construct

Cited 1 time

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The pCaspeR4-Tubp-miniCic-linker-mScarlet-I was obtained by amplifying by PCR miniCic-linker from pCaspeR4-Tubp-miniCic-linker-mCherry(Moreno et al., 2019 (link)) and mScarlet-I from pmScarlet-i_C1 (Addgene 85044). These two inserts were cloned in the vector pCaspeR4-TubP-Gal80 linearised by NotI, XbaI digestion (to excise Gal80) using NEBuilder HiFi DNA Assembly Method. The construct was checked by sequencing and inserted through P-element after injection by Bestgene. Details about the primers used for this construct can be found in the key resources table.
The construct was tested by comparing the dynamics and pattern in the notum compared to previously characterised dynamics using miniCic-mCherry (Moreno et al., 2019 (link)). Similar dynamics were also observed at the single cell level between miniCic-mScarlet and the FRET sensor ubi-EKARnls (Ogura et al., 2018 (link)) (Figure S4).

Valon L., Davidović A., Levillayer F., Villars A., Chouly M., Cerqueira-Campos F, & Levayer R. (2021). Robustness of epithelial sealing is an emerging property of local ERK feedback driven by cell elimination. Developmental Cell, 56(12), 1700-1711.e8.

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Cloning LOTR1 Promoter and Coding Sequence

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A 3 kb promoter fragment of LOTR1 was generated by 5’-aacaggtctcgacctctttctatctgtttgtacctaagt-3’ and 5’-aacaggtctcatgttaactagagaatgtacggcttgttt-3’ and cloned via Eco31I-restriction sites into GreenGate entry module vector pGGA000 (Lampropoulos et al., 2013 (link)). Genomic fragment of LOTR1 coding sequence was amplified with 5’-aacaggtctcaggctcgatgTCAGCTATTCATCTTAAAAACCAAACTTCA and 5’-aacaggtctcaCTGAAGGACACCTTGGGTTCCG-3’ and cloned into pGGC000. mScarlet-I was amplified from pmScarlet-I_C1 (Bindels et al., 2017 (link)) obtained from Addgene and secretion peptide was added by tandem PCR amplification with 5’-gctctttccctctatctcctgcccaatccagccactagtATGGTGAGCAAGGGCGAG-3’ (FW1) and 5’-aacaGGTCTCaAACAatgaaagccttcacactcgctctcttcttagctctttccctctatctcctg-3’ (FW2) and 5’-aacaGGTCTCaAGCCCTTGTACAGCTCGTCCATGC-3’ (RV) and cloned via Eco31I sites into pGGB000. Final destination clones were obtained by subsequent GoldenGate reaction with pGreenII based pGGZ003 and transformed into Arabidopsis using floral dip.

Kolbeck A., Marhavý P., De Bellis D., Li B., Kamiya T., Fujiwara T., Kalmbach L, & Geldner N. (2022). CASP microdomain formation requires cross cell wall stabilization of domains and non-cell autonomous action of LOTR1. eLife, 11, e69602.

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