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Pcag cfp

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The PCAG-CFP is a plasmid vector designed for the expression of enhanced cyan fluorescent protein (CFP) in mammalian cells. It contains the necessary elements for stable transfection and expression of the CFP reporter gene.

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Lab products found in correlation

Pcag dsred, by Addgene (3 mentions) Pcag yfp, by Addgene (3 mentions) Pcag gfp, by Addgene (2 mentions) Pcalnl dsred, by Addgene (2 mentions) Prl tk, by Promega (2 mentions) Pcag egfp, by Addgene (1 mentions)

5 protocols using pcag cfp

1

Plasmid-based CRISPR-Cas9/Cpf1 Expression

Cited 2 times
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A list and partial sequences of plasmids used in this study can be found in Supplementary Note 1; crRNA sequences are listed in Supplementary Table 1 and oligonucleotide sequences are found in Supplementary Table 5. AsCpf1 and LbCpf1 human expression plasmids (SQT1659 and SQT1665, respectively) were generated by inserting the open-reading frames of these nucleases from plasmids pY010 and pY016 (Addgene plasmids # 69982 and # 69988, respectively) into the NotI and AgeI sites of pCAG-CFP (Addgene # 11179). Plasmid SQT817 was used to express SpCas9 (ref 28 (link)). Oligonucleotide duplexes corresponding to spacer sequences were annealed and ligated into BsmBI-digested BPK3079, BPK3082, and BPK1520 (ref 24 (link)) for U6 promoter-driven expression of AsCpf1, LbCpf1, and SpCas9 gRNAs, respectively. New plasmids described in this study will be deposited with the non-profit plasmid repository Addgene: http://www.addgene.org/crispr-cas.
Kleinstiver B.P., Tsai S.Q., Prew M.S., Nguyen N.T., Welch M.M., Lopez J.M., McCaw Z.R., Aryee M.J, & Joung J.K. (2016). Genome-wide specificities of CRISPR-Cas Cpf1 nucleases in human cells. Nature biotechnology, 34(8), 869-874.
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2

Diverse Fluorescent Protein Expression Vectors

Cited 1 time
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pCAG-GFP (Addgene plasmid 11150) 39 (link), pCAG-YFP (Addgene plasmid 11180) 39 (link), pCAG-CFP (Addgene plasmid 11179) 39 (link), pCAG-tdT (Cepko lab, Harvard Medical School), pCAG-mCherry (Cepko lab, Harvard Medical School), pCAG-DsRed (Addgene plasmid 11151) 39 (link), pRL-TK (Promega, #E2241), pRho-GFP-IRES-AP (referred to as Rho-GFP in main text) 40 (link), pCAG-nlacZ (Cepko lab, Harvard Medical School). pAAV-CAG-FLEX-tdT was a gift from Edward Boyden (Addgene plasmid #28306), pCALNL-DsRed (Addgene plasmid #13769) 41 (link).
Tang J.C., Rudolph S., Dhande O.S., Abraira V.E., Choi S., Lapan S., Drew I.R., Drokhlyansky E., Huberman A.D., Regehr W.G, & Cepko C.L. (2015). Cell Type-Specific Manipulation with GFP-Dependent Cre Recombinase. Nature neuroscience, 18(9), 1334-1341.
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3

Fluorescent Plasmid Validation Protocol

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Plasmids encoding fluorescent reporter proteins were used to validate electroporation: pCAG‐DsRed (Addgene: 11151), pCAG‐YFP (Addgene: 11180), pCAG‐EGFP (Addgene: 11150), pCAG‐CFP (Addgene: 11179), pCBA‐TdTomato (Addgene: 28017). Plasmids were amplified and purified according to the suppliers’ recommendations, filtered, and stored at −20°C until use.
Dempsey B., Turner A.J., Le S., Sun Q., Bou Farah L., Allen A.M., Goodchild A.K, & McMullan S. (2015). Recording, labeling, and transfection of single neurons in deep brain structures. Physiological Reports, 3(1), e12246.
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4

Diverse Fluorescent Protein Expression Vectors

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
pCAG-GFP (Addgene plasmid 11150) 39 (link), pCAG-YFP (Addgene plasmid 11180) 39 (link), pCAG-CFP (Addgene plasmid 11179) 39 (link), pCAG-tdT (Cepko lab, Harvard Medical School), pCAG-mCherry (Cepko lab, Harvard Medical School), pCAG-DsRed (Addgene plasmid 11151) 39 (link), pRL-TK (Promega, #E2241), pRho-GFP-IRES-AP (referred to as Rho-GFP in main text) 40 (link), pCAG-nlacZ (Cepko lab, Harvard Medical School). pAAV-CAG-FLEX-tdT was a gift from Edward Boyden (Addgene plasmid #28306), pCALNL-DsRed (Addgene plasmid #13769) 41 (link).
Tang J.C., Rudolph S., Dhande O.S., Abraira V.E., Choi S., Lapan S., Drew I.R., Drokhlyansky E., Huberman A.D., Regehr W.G, & Cepko C.L. (2015). Cell Type-Specific Manipulation with GFP-Dependent Cre Recombinase. Nature neuroscience, 18(9), 1334-1341.
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5

Plasmid-based CRISPR-Cas9/Cpf1 Expression

Cited 2 times
Check if the same lab product or an alternative is used in the 5 most similar protocols
A list and partial sequences of plasmids used in this study can be found in Supplementary Note 1; crRNA sequences are listed in Supplementary Table 1 and oligonucleotide sequences are found in Supplementary Table 5. AsCpf1 and LbCpf1 human expression plasmids (SQT1659 and SQT1665, respectively) were generated by inserting the open-reading frames of these nucleases from plasmids pY010 and pY016 (Addgene plasmids # 69982 and # 69988, respectively) into the NotI and AgeI sites of pCAG-CFP (Addgene # 11179). Plasmid SQT817 was used to express SpCas9 (ref 28 (link)). Oligonucleotide duplexes corresponding to spacer sequences were annealed and ligated into BsmBI-digested BPK3079, BPK3082, and BPK1520 (ref 24 (link)) for U6 promoter-driven expression of AsCpf1, LbCpf1, and SpCas9 gRNAs, respectively. New plasmids described in this study will be deposited with the non-profit plasmid repository Addgene: http://www.addgene.org/crispr-cas.
Kleinstiver B.P., Tsai S.Q., Prew M.S., Nguyen N.T., Welch M.M., Lopez J.M., McCaw Z.R., Aryee M.J, & Joung J.K. (2016). Genome-wide specificities of CRISPR-Cas Cpf1 nucleases in human cells. Nature biotechnology, 34(8), 869-874.
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