Table 3.
Cas13-based RNA technologies.
| Application Field | Cas Effectors | Efficiency | Application |
|---|---|---|---|
| Basic biochemical research | Cas13a | Medium | RNA knockdown, Nucleic acid detection [46] |
| Cas13a | 2 × 103 copies/mL | Nucleic acid detection (SHERLOCK) [47] | |
| Cas13a | High (90%) | Virus interference, transcript targeting guide-induced, gene silencing [48] | |
| Cas13d | High (89%) | Chemically modified crRNAs can modify the transcriptome of human primary T cells [49] | |
| Cas13a | 40.4–83.9% | Demonstrates the RNA-guided RNase activity of the Cas13a [46] | |
| Cas13b | Discovery and biochemical activity of Cas13b [43] | ||
| Cas13a, Cas13b |
RNA interference, virus interference, and virus resistance [50] | ||
| Cas13d | 34–46% | Therapeutic potential, generation of AAV all-in-one vector consisting of up to three pre-sgRNAs for effective knockdown of VEGFA gene expression [45] | |
| Cas13d | Discovery and biochemical activity of Cas13d [36] | ||
| Cas13X | Discovered Cas13x effector; exhibited robust editing efficiency and high specificity to induce RNA base conversions [13] | ||
| Nucleic acid detection | Cas13a | Medium | Nucleic acid detection (SHERLOCKv2) [47] |
| Cas13a | High | One-step experimental screening system, diagnostics, and therapeutics for COVID-19 [51] | |
| Cas13a | High (100%) | RT-LAMP, point-of-care diagnostics, detect SARS-CoV-2 [52] | |
| Cas13a | CRISPR diagnostics and targeted cancer therapy [53] | ||
| Cas13a | High (90%) | Lateral flow strip, single-step SARS-CoV-2, two-step SARS-CoV-2 assay (SHINE) [54] | |
| Cas13a | Hepatitis B virus covalently closed circular DNA detection [55] | ||
| Nucleic acid imaging | Cas13d | CRISPR Live-cell fluorescent in situ hybridization (LiveFISH) accurately detects chromosomal disorders and tracks the real-time movement of DNA double-strand breaks [56,57] | |
| Cas13b | Dynamic imaging of RNA in living cells, simultaneous visualization of RNA-RNA and DNA-RNA in living cells [58] | ||
| Cas13b | RNA-protein interactions identify proteins associated with an endogenous RNA, CRISPR-based RNA proximity proteomics (CBRPP) [59] | ||
| Cas13a | Endogenous RNA foci imaging of RNA in the nucleus and cytoplasm in living cells [60] | ||
| Antiviral application | Cas13d | 90% | CRISPR-based strategy for RNA-guided viral RNA inhibition and degradation (PAC-MAN) [61] |
| Cas13b | 50% | Abrogation of pRRSV infectivity in mammalian cells [62] | |
| Cas13a | RNA virus resistance in both dicot and monocot plants [63] | ||
| Cas13a | 70–84% | CRISPR-Cas13a mediated targeting of hepatitis C virus internal-ribosomal entry site (IRES) [64] | |
| Disease treatment | Cas13d | Using pgRNAs can robustly suppress the propagation of plant RNA viruses [65] | |
| Cas13a | GIGS offers a novel and flexible approach to RNA reduction for crop improvement and functional genomics [48] | ||
| Cas13X | Rescue of autosomal dominant hearing loss by in vivo delivery [66] | ||
| Cas13b | RNA base editing; an efficient RNA base editor, dPspCas13b-RESCUE-NES, a potentially useful tool for biomedical research and genetic disease [67] | ||
| Cas13a, Cas13b, Cas13d |
A one-step platform for screening high-efficient and minimal off-target CRISPR/Cas13 crRNAs to eradicate the SARS-CoV-2 virus for the treatment of COVID-19 patients [51] | ||
| Cas13a | Detects BK polyomavirus DNA and cytomegalovirus DNA from patient-derived blood and urine samples [68] | ||
| Cas13d | 100% | PiggyBac systems; Cas13d vector achieved extremely high efficiency in RNA knockdown (98% knockdown for CD90) with optimized gRNA designs [69] | |
| Cas13d | 99% | Alleviation of neurological disease by RNA editing [70] | |
| Cas13d | Establish RfxCas13d as a versatile platform for knocking down gene expression in the nervous system [71] | ||
| Cas13 | 50% | A versatile tool for cancer diagnosis, therapy, and research [72] | |
| Cas13 | Cas13s for targeting viral RNA [53] |