Optimization of a Cas12a-Driven Synthetic Gene Regulatory Network System.
Kang H, Fitch JC, Varghese RP, Thorne CA, Cusanovich DA
ACS Synth Biol. 2025 May 16;14(5):1732-1744. doi: 10.1021/acssynbio.5c00084. Epub 2025 May 2.
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Plasmids from Article
ID | Plasmid | Purpose |
---|---|---|
202041 | EF1a-Triplex-28-M13-28-pA | Encodes a non-coding transcript expressed from the EF1a promoter that has a 3' UTR containing a MALAT1 triplex, Csy4 binding sites ("28"), and a cloning site for a Cas9 gRNA. |
202042 | pCE059-SiT-ddCas12a(E993A)-1xNLS-[Activ]-triplex-DR-M13-DR-pA | Encodes a DNase-dead AsCpf1 fused to a NLP nuclear localization signal (NLS) and [Activ] transactivation domain. 3' UTR has a MALAT1 triplex, dCas12a direct repeats (DR), and a crRNA cloning site. |
202043 | pCE059-SiT-ddCas12a(E993A)-3xNLS-[Activ]-triplex-DR-M13-DR-pA | Encodes a DNase-dead AsCpf1 fused to a 3x SV40 nuclear localization signal (NLS) and [Activ] transactivation domain. 3' UTR has a MALAT1 triplex, dCas12a direct repeats (DR), and a crRNA cloning site. |
202044 | pCE059-SiT-ddCas12a(E993A)-3xNLS-VP64-triplex-DR-M13-DR-pA | Encodes a DNase-dead AsCpf1 fused to a 3x SV40 nuclear localization signal (NLS) and VP64 transactivation domain. 3' UTR has a MALAT1 triplex, dCas12a direct repeats (DR), and a crRNA cloning site. |
202045 | Px-PhiYFP-pA | Encodes PhiYFP, which can be expressed with the activation of a synthetic promoter by Cas enzymes. A cloning site is provided to construct the synthetic Cas-targeted promoter upstream of PhiYFP. |
202046 | Px-PhiYFP-triplex-DR-M13-DR-pA | Encodes PhiYFP downstream of a cloning site for constructing a synthetic Cas-targeted promoter. 3’ UTR has a MALAT1 triplex, dCas12a DRs, and cloning site for crRNA to activate downstream targets. |
202047 | Px-PhiYFP-triplex-28-M13-28-pA | Encodes PhiYFP downstream of a cloning site for constructing a synthetic Cas-targeted promoter. 3’ UTR has a MALAT1 triplex, Csy4 28nt sites, and cloning site for gRNA to activate downstream targets. |
202048 | Px-mCerulean-pA | Encodes mCerulean, which can be expressed with the activation of a synthetic promoter by Cas enzymes. A cloning site is provided to construct the synthetic Cas-targeted promoter upstream of mCerulean. |
202049 | Px-mCherry-pA | Encodes mCherry, which can be expressed with the activation of a synthetic promoter by Cas enzymes. A cloning site is provided to construct the synthetic Cas-targeted promoter upstream of mCherry. |