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M st1n vp64

Manufactured by Addgene
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The M-ST1n-VP64 is a lab equipment product. It is a fusion protein consisting of the CRISPR-associated protein ST1 Cas9 nuclease and the VP64 transcriptional activator domain. This product is designed for gene regulation applications.

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Lab products found in correlation

Cas9m4 vp64, by Addgene (2 mentions)

2 protocols using m st1n vp64

1

Activation Domain Cloning and dCas9 Constructs

Cited 2 times
Check if the same lab product or an alternative is used in the 5 most similar protocols
Activation domains were cloned using a combination of Gibson and Gateway assembly or Golden Gate assembly methods. For experiments involving multiple activation domains, ADs were separated by short glycine-serine linkers. Activator sequences are listed in the Supplementary Data (vectors to be deposited in Addgene). All SP-dCas9 plasmids were based on Cas9m4-VP64 (Addgene #47319)6 (link), ST1-dCas9 plasmids were based on M-ST1n-VP64 (Addgene #48675)15 (link). Sequences for gRNAs are listed in the supplementary information. gRNAs for endogenous human gene activation were selected to bind between 1 and 1000 bp upstream of the transcriptional start site (TSS). gRNAs for iPSC differentiation to iNeurons, targeting NGN2 and NEUROD1, were selected to bind between 1 and 2000 base pairs upstream of the transcriptional start site. All human gRNAs were expressed from either cloned plasmids (Addgene #41817)5 (link) or integrated into the genome through lentiviral delivery (plasmid SB700). Guide RNA sequences are listed within the Supplementary Data. Reporter targeting gRNAs were previously described (Addgene #48671 and #48672)6 (link).
Chavez A., Scheiman J., Vora S., Pruitt B.W., Tuttle M., Iyer E., Lin S., Kiani S., Guzman C.D., Wiegand D.J., Ter-Ovanesyan D., Braff J.L., Davidsohn N., Housden B.E., Perrimon N., Weiss R., Aach J., Collins J.J, & Church G.M. (2015). Highly-efficient Cas9-mediated transcriptional programming. Nature methods, 12(4), 326-328.
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2

Activation Domain Cloning and dCas9 Constructs

Cited 2 times
Check if the same lab product or an alternative is used in the 5 most similar protocols
Activation domains were cloned using a combination of Gibson and Gateway assembly or Golden Gate assembly methods. For experiments involving multiple activation domains, ADs were separated by short glycine-serine linkers. Activator sequences are listed in the Supplementary Data (vectors to be deposited in Addgene). All SP-dCas9 plasmids were based on Cas9m4-VP64 (Addgene #47319)6 (link), ST1-dCas9 plasmids were based on M-ST1n-VP64 (Addgene #48675)15 (link). Sequences for gRNAs are listed in the supplementary information. gRNAs for endogenous human gene activation were selected to bind between 1 and 1000 bp upstream of the transcriptional start site (TSS). gRNAs for iPSC differentiation to iNeurons, targeting NGN2 and NEUROD1, were selected to bind between 1 and 2000 base pairs upstream of the transcriptional start site. All human gRNAs were expressed from either cloned plasmids (Addgene #41817)5 (link) or integrated into the genome through lentiviral delivery (plasmid SB700). Guide RNA sequences are listed within the Supplementary Data. Reporter targeting gRNAs were previously described (Addgene #48671 and #48672)6 (link).
Chavez A., Scheiman J., Vora S., Pruitt B.W., Tuttle M., Iyer E., Lin S., Kiani S., Guzman C.D., Wiegand D.J., Ter-Ovanesyan D., Braff J.L., Davidsohn N., Housden B.E., Perrimon N., Weiss R., Aach J., Collins J.J, & Church G.M. (2015). Highly-efficient Cas9-mediated transcriptional programming. Nature methods, 12(4), 326-328.
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