Toolkit Items

The AS1411 aptamer-modified cell membrane biomimetic core-shell system is a light-augmented delivery harness for CRISPR-Cas9 plasmid cargo (pCas9). It consists of a cell membrane-camouflaged shell modified with AS1411 aptamers and photosensitizers to promote tumor targeting, reactive oxygen species-mediated lysosomal escape, and light-controllable pCas9 release for enhanced gene editing.

Caged guide RNAs are synthetic CRISPR guide RNAs containing photolabile nucleobase substitutions in the 5′ protospacer that enable light-activated control of Cas9 function. They were developed to conditionally regulate genome editing in mammalian cells and zebrafish embryos by suppressing guide RNA binding to target DNA until optical activation.

ChemHEAL is a small-molecule-inducible variant of the HEAL dCas12f-based CRISPR activation platform for programmable transcriptional control. It uses a multi-component architecture in which transcriptional activators are recruited through MS2 coat protein binding to MS2 aptamers embedded in the sgRNA scaffold.

HEAL is a compact, inducible, multi-component dCas12f-based CRISPR activation system for programmable transcriptional activation in vivo. It uses sgRNA-embedded MS2 aptamers to recruit transactivators through MS2 coat protein binding at CRISPR-targeted DNA sites.

The hypoxanthine switch is a small molecule-responsive RNA element described as a hypoxanthine-sensitive switch. Available evidence indicates that its intracellular switch activity correlates with aptamer binding properties measured biochemically.

The light-controlled CRISPR/dCas9 transactivation system is a multi-component optogenetic transcriptional activator that couples CRISPR/dCas9-mediated gene activation to a light-responsive PAL–RNA aptamer interaction. It enables reversible optical control of transactivation and was described as a variation of the CRISPR/dCas9 system for light-controlled activation of gene expression.

mOptoT7 is a mammalian optogenetic transcription system composed of a split T7 RNA polymerase fused to the blue-light-inducible nMag/pMag Magnets photodimerization system. Blue light drives reconstitution of the split polymerase to activate transcription from orthogonal T7 promoters in mammalian cells, and the system has been used to produce protein-coding mRNA, shRNA, and the Pepper RNA aptamer.

OptoHEAL is described as a red-light-inducible version of the HEAL CRISPR activation platform for remote and precise transcriptional control. HEAL is a compact dCas12f-based CRISPRa architecture that activates transcription by recruiting transactivators through MS2 coat protein binding to MS2 aptamers embedded in the sgRNA scaffold.

This tool is an RNA aptamer-based component for light-controlled, reversible gene transactivation in a CRISPR/dCas9-based system. It is built on the interaction between the photoreceptor PAL and an RNA aptamer to regulate gene expression with light.

SIBR-Cas, termed Self-splicing Intron-Based Riboswitch-Cas, is a multi-component bacterial CRISPR genome engineering system that provides inducible control over CRISPR-Cas counterselection. It is reported to delay counterselection to permit editing events and has been applied to gene knockout in bacteria with poor homologous recombination systems.

Stem-loop prime editors (sPEs) are modified prime editing guide RNA constructs in which stem-loop aptamers are added to the 3′ terminus of the pegRNA. This altered pegRNA architecture is used as a basis for tethered prime editors and for split pegRNA prime editor configurations.

Tethered prime editors (tPEs) are prime editing constructs in which stem-loop aptamer-modified pegRNAs are tethered to Cas9 nickase. The design alters pegRNA architecture to increase prime editing efficiency and flexibility.