Table 2 Targeting miRNA genes and miRNA transcripts by CRISPR/Cas in vivo and in vitro studies
From: Targeting miRNA by CRISPR/Cas in cancer: advantages and challenges
Type of CRISPR | Purpose of use | miRNA | Target | Type of diseases | Cell line (in vitro) | Animal (in vivo) | Vector | Finding | References |
|---|---|---|---|---|---|---|---|---|---|
CRISPR/dCas9 | Therapy | miR-155 | – | Liver cancer | HEK293T, AML12 | – | Lentivirus | Exosomes are modified to transport and deliver novel RNA payload to target cells | [84] |
CRISPR/Cas9 | Therapy | miR-155 | S PU.1, AID, SHIP1, SMAD5, and SOCS1 | RA | RAW264.7 and HEK293T | – | Lentivirus | Genome editing with miRNA-155 holds promise as a treatment for RA | [82] |
CRISPR/Cas9 | Function investigation | 10 miRNA genes | – | – | – | Zebrafish | – | Cas9 nuclease with 2, 4, 10 or > 24 multiplexed sgRNA can cause mutations in 90% of miRNA genome | [85] |
CRISPR/Cas9 | Therapy and pathway investigation | miRNA-26a-1 miRNA-26a-2 | – | Respiratory distress syndrome | – | Mice (C57BL/6J and FVB) | – | miR-26a has a role in PS synthesis in AECIIs | [86] |
CRISPR/Cas9 | Function investigation | miR-196a miR-219 | – | Neurocristopathies | – | Xenopus | – | miRNA mimics have been applied to recover the knockout phenotype Knocking out miR-219 and miRNA-106a related to loss of NC and hatching gland abnormalities in mice | [87] |
CRISPR/Cas9 | Therapy | miR122 | – | HCV | Huh7 | – | Adeno-associated virus | Creating tailored cellular clones that resemble to the parental cells yet are immune to HCV multiplication and infection | [88] |
CRISPR/Cas9 | Function investigation | miR-4018a | – | – | – | Ciona embryo | Electroporation | MICR-ON is used to observe and analyze miRNA expression and function in a living organism and its biological system | [89] |
CRISPR/Cas9 | Function investigation | miR17 family | Fog2 | – | E14 | Mouse | – | Varied members of the miRNA17 family (14 miRNAs) have different functions in embryonic stem cell development | [90] |
CRISPR/Cas9 | Investigation | – | – | – | HEK293T | – | – | A pool of transiently transfected cells must allow functional examination of a hypothesized miRNA-target combination using clonal cell lines or transgenic animals | [91] |
CRISPR/Cas9 | Therapy | miR-17 miR-200c miR-141 | – | Colorectal cancer | HCT116 and HT-29 | – | Lenti-CRISPR | Suppressing miRNAs by up to 96% in robustness by using CRISPR/Cas9 CRISPR/Cas9 regulates off-targeting on miRNAs from the same family or with a similar sequence It has been demonstrated how CRISPR/Cas9 miRNA knockdown is stable over the long term | |
CRISPR/Cas9 | Testing the methods | miRNA-29b1 | – | – | – | Mice (C57BL/6) | – | Knocking out miRNA-29b1 gene in mice A 10 bp deletion, a 23 bp loss, and a 3 bp insertion have been seen in mouse genotypes miRNA-29b1 expression was down-regulated in the kidneys, liver, heart, spleen, and lung | [93] |
CRISPR/Cas9 | Function investigation | miR-31-5p miR-92b-3p miR-146b-5p miR-151a-3p miR-194-5p miR-95-3p miR-181a-5p miR-188-5p miR-196b-5p miR-584-5p miR-1304-3p miR-100-5p miR-149-5p | – | Cervical and gastric cancer | HeLa or NCI-N87 | – | Lentivirus | Five HeLa pro-fitness and cervical cancer up-regulated miRNAs were found There was an up-regulation of six NCI-N87 profit and gastric cancer miRNAs Three down-regulated and anti-fitness miRNAs were found | [94] |
CRISPR/Cas9 | Function investigation | miR-497 miR-195 miR-143 miR-145 | – | Cardiovascular diseases and cancer | VSMCs, HEK293T | – | Lentivirus | Editing miR-195 decreased miR-497a expression in the miR-497195 cluster. Despite the absence of gene editing in the miR-497a genomic region, computational simulation demonstrated a change in the three-dimensional form of the pri-miR-497-195 | [95] |