Pcag vector

Manufactured by Addgene

The PCAG vector is a plasmid that can be used for gene expression in mammalian cell lines. It contains a constitutive promoter and a multiple cloning site for inserting the gene of interest.

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Lab products found in correlation

Fetal calf serum (fcs), by Thermo Fisher Scientific (1 mentions) Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (1 mentions) Block it pol 2 mir rnai expression vector kit, by Thermo Fisher Scientific (1 mentions) Block it rnai designer tool, by Thermo Fisher Scientific (1 mentions) Mir ctrl, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

PCAG-GFP vector (6 citations) PCAG-Myc vector (6 citations) PCAG-mCherry-gRNA vector (5 citations) PCAG-hCas9 vector (3 citations) PCAG-EGxxFP vector (2 citations) PCAG-Cre:GFP vector (2 citations) PCAG-mCherry-sgRNA vector (2 citations)

2 protocols using pcag vector

Characterizing SRSF1 Mutant Variants

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Human embryonic kidney HEK293 cells were maintained in Dulbecco’s modified Eagle’s medium (GibcoBRL) supplemented with 10% fetal calf serum (GibcoBRL). The mammalian vectors expressing FLAG-tagged SRSF1 WT, Y19A, and E87N were obtained by cloning the SRSF1 ORF, amplified by PCR, in frame into the pCAG vector (Addgene). SRSF1 Y19A and E87N mutants were created by site-directed mutagenesis using specific primers.

Cléry A., Krepl M., Nguyen C.K., Moursy A., Jorjani H., Katsantoni M., Okoniewski M., Mittal N., Zavolan M., Sponer J, & Allain F.H. (2021). Structure of SRSF1 RRM1 bound to RNA reveals an unexpected bimodal mode of interaction and explains its involvement in SMN1 exon7 splicing. Nature Communications, 12, 428.

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Neuronal miR-Tlr8 Expression Construct

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For the miR-Tlr8 construct, the oligonucleotide sequence 5′-TGCTGTTTCAAACCAGGTAGAAGGAAGTTTTGGCCACTGACTGACTTCCTTCTCTGGTTTGAAA-3′ designed by the BLOCK-iT RNAi Designer tool (Invitrogen) was used. The paired oligonucleotides were inserted into a pcDNA6.2-GW/EmGFPmiR vector using the BLOCK-iT Pol II miR RNAi Expression Vector kit (Invitrogen). The plasmid pcDNA6.2-GW/EmGFP-miR-neg (miR-Ctrl; Invitrogen), predicted to not target any vertebrate gene, was used as a negative control. The negative control sequence is 5′-GAAATGTACTGCGCGTGGAGACGTTTTGGCCACTGACTGACGTCTCCACGCAGTACATTT-3′. These plasmids in the pcDNA vector were used in neuronal cultures. For IUE, the miRNA fragment in pcDNA 6.2-GW/EmGFP-miR-neg and -Tlr8 was further subcloned into the 3′ untranslated region of the GFP in pCAG vector (11150; Addgene; provided by C. Cepko; Matsuda and Cepko, 2004 (link)).

Hung Y.F., Chen C.Y., Shih Y.C., Liu H.Y., Huang C.M, & Hsueh Y.P. (2018). Endosomal TLR3, TLR7, and TLR8 control neuronal morphology through different transcriptional programs. The Journal of Cell Biology, 217(8), 2727-2742.

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