Skip to main content
Addgene

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. (Link opens in a new window) PubMed (Link opens in a new window) Article

Plasmids from Article

ID Plasmid Purpose
202041EF1a-Triplex-28-M13-28-pAEncodes 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.
202042pCE059-SiT-ddCas12a(E993A)-1xNLS-[Activ]-triplex-DR-M13-DR-pAEncodes 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.
202043pCE059-SiT-ddCas12a(E993A)-3xNLS-[Activ]-triplex-DR-M13-DR-pAEncodes 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.
202044pCE059-SiT-ddCas12a(E993A)-3xNLS-VP64-triplex-DR-M13-DR-pAEncodes 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.
202045Px-PhiYFP-pAEncodes 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.
202046Px-PhiYFP-triplex-DR-M13-DR-pAEncodes 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.
202047Px-PhiYFP-triplex-28-M13-28-pAEncodes 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.
202048Px-mCerulean-pAEncodes 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.
202049Px-mCherry-pAEncodes 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.

Antibodies from Article