Applications of CRISPR-Cas9 as an Advanced Genome Editing System in Life Sciences
Abstract
:1. Introduction
2. Origin, Development, and Mechanism of the CRISPR-Cas9 System
3. Ethical Issues in Genome Editing by CRISPR-Cas9 System
4. Applications of CRISPER-Cas9 Technology
4.1. Human Science
4.1.1. Clinical Trials Using CRISPR-Cas9 Technology
4.1.2. Clinical Trials of the Eye Based on CRISPR-Cas9
4.1.3. Limitations of CRISPR-Cas-Based Gene Therapy
4.2. Plant Science
4.2.1. Plant Disease Resistance
4.2.2. Yield of Crop Plants
4.2.3. Genome Modification for Nutritional Improvement
4.2.4. Medicinal Plants
4.3. Animal Breeding
4.3.1. Modification of Pigs for Xenotransplantation Research
4.3.2. Application of CRISPR-Cas9 Technology in Insects
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Feature | CRISPR-Cas | TALEN | ZFN |
---|---|---|---|
Cost | Low | High | Low |
Ease of design | Simple | A little complex | Moderate |
Specificity | High | Intermediate | Low |
Pros | Modifies multiple sites in tandem | Highly effective and specific | Highly effective and specific |
Cons | PAM motif required next to target sequence | Time consuming | Time consuming |
Multiplex genome editing | High-yield multiplexing | Few models | Few models |
Organism | Risks | Bioethical Issues | References |
---|---|---|---|
Bacteria | Gene mutations/ Gene drifts | Disruption of ecological balance | [20,24,25] |
Plants | Gene mutations/ Gene drifts | Disruption of ecological balance | [20,26] |
Animals/ chimeric animals | Gene mutations | Disruption of ecological balance | [24,27,28,29,30,31,32] |
Humans | Gene mutations Side effects Cost Genetic mosaicism | Eugenics Informed consent Enhancement Accessibility Patenting Safety Incomplete or over legislation | [21,24,26,33,34,35,36,37] |
Type | Oncogene | Tumor Suppressor Gene | Drug-Resistance Gene |
---|---|---|---|
Breast | SHCBP1, MIEN1, miR-27b, mi23b, HER2 exons | CPEB2, ETS1, BRCA1 | HER2, EGFR, ER |
Prostate | PCAR19, CECAM21, SENP1, miR-302/367, miR-1205 | PGC1a, DEPTOR, p53, RB1 | BLS-211, NANOGP8, NANOG1 |
Lung | IGFIR, ERBB2, RSF1, FOS, MCM4 | MFN2, MiR-1205, GOT1, TP53 | NRF2, MiR-1205, ER300, RSF1 |
Liver | Plxnb1, NCAPG, CDK7, IncBRM, Nf1 | BAP1, HELLS, Tp53, Traf3 | NF1, MED12, ERK2 |
Colorectal | KRAS, HPV16, Fut4, NRAS | LIMCH, PTEN, SOX15 | miR-139-5P, ZEB1 |
CRISPR-Cas9 function | Knockout | Activate | Promote drug sensitivity |
Target Gene and Effect | Disease | Intervention |
---|---|---|
Cas9-mediated creation of CD19 and CD20 | Leukemia | CAR T cells to CD19 and CD20 or CD19 and CD22 |
CCR5 knockout | HIV | Modified CD34+ hematopoietic stem cells |
CD7 knockout in CD7 CAR T cells | T-cell malignancies | CAR T cells to CD7 and knockout of native CD7 to prevent self-targeting |
Correction of the hemoglobulin subunit β globulin gene | β-thalassemia | Ex vivo modified hematopoietic stem cells |
Creation of a CD19-directed T cell | Refractory B-cell malignancies | CD19-directed T-cell immunotherapy |
Cytokine-induced SH2 protein (CISH) knockout | Metastatic gastrointestinal epithelial cancer | Modified tumor-infiltrating lymphocytes |
disruption of HPK1 | Refractory B cell malignancies | CD19-CAR modified T cells with CAR delivered by lentivirus and Cas9 knockout of HPK1 |
Disruption of the erythroid enhancer to BCL11A gene | β-thalassemia | Ex vivo modified hematopoietic stem cells |
Sickle cell anemia | ||
β-thalassemia and severe sickle cell anemia | Ex vivo- modified hematopoietic stem cells, 15-year follow-up study | |
E6 and E7 oncogene of HPV16 and HPV18 deletion | HPV-related malignancy | Plasmid in a gel containing a polymer to facilitate delivery |
Programmed cell death protein 1 (PD-1) knockout | Mesothelin positive solid tumors | CAR T cells to mesothelin with added PD-1 and TCR knockout |
Hormone refractory prostate cancer | Modified T cells | |
Esophageal cancer | ||
Metastatic non-small cell lung cancer | ||
Stage IV bladder cancer | ||
Metastatic renal cell carcinoma | ||
EBV-positive, advanced stage malignancies | Modified T cells selected for those targeting EBV positive cells | |
Mesothelin positive solid tumors | CAR T cells to mesothelin with PD-1 knockout | |
Removal of alternative splice site in CEP290 | Leber congenital amaurosis 10 | ZFN-mediated removal of intronic alternative splice site in retinal cells |
TCRα, TCRβ, PD-1 knockout | Various malignancies | Modified T cells with Cas9-mediated deletions and lentiviral transduction of NY-ESO-1 targeted TCR |
βTCRα, TCRβ, β-2 microglobin (B2M) knockout | B-cell leukemia | CD19-CAR modified T cells with CAR delivered by lentivirus and Cas9 knockout B2M and TCR to create universal T cells |
Virus | Type of Nucleic Acid | Involved Protein | Plant under Attack | References |
---|---|---|---|---|
Beet severe curly top virus | DNA | Cas9 | Capsicum | [75] |
Bean yellow dwarf virus | DNA | Cas9 | Oat | [76] |
Turnip mosaic virus | RNA | Cas13 | Cruciferous plants, Chinese cabbage, turnip, mustard, radish | [77] |
Tomato yellow leaf curl virus | DNA | Cas9 | Invading a number of seeds, including tomato | [78,79] |
Yellowing virus | RNA | Cas13 | Cucumber | [80] |
Zucchini yellow mosaic virus | RNA | Cas13 | Cucumber | [81] |
Papaya ring spot mosaic virus | RNA | Cas13 | Cucumber | [82] |
Plant | Targeted Area in Gene | Disease | References |
---|---|---|---|
Rice | Mutagenesis of the ERF Transcription Factor Gene OsERF922 | Blast | [72] |
Duncan grapefruit | Effector-binding element in the promoter of the Lateral Organ Boundaries 1 gene | Citrus bacterial canker (CBC) | [81] |
Wanjinchen oranges | (CsLOB1G and CsLOB1−) alleles | Citrus bacterial canker (CBC) | [82] |
Crop | Method | Target Gene | Stress/Trait | References |
---|---|---|---|---|
A. thaliana/ N. benthamiana | NHEJ | dsDNA of virus (A7, B7, and C3 regions) | Beet severe curly top virus resistance | [75] |
N. benthamiana Bean | NHEJ | BeYDV | Yellow dwarf virus (BeYDV) resistance | [76] |
N. benthamiana | NHEJ | ORFs and the IR sequence sDNA of virus | Tomato yellow leaf curl virus (TYLCV) and Merremia mosaic virus (MeMV) | [77] |
Rice | NHEJ | OsERF922 (ethylene responsive factor) | Blast Resistance | [78] |
Cucumber | NHEJ | eIF4E (eukaryotic translation initiation factor 4E) | Cucumber vein yellowing virus (CVYV), Zucchini yellow mosaic virus (ZYMV), and (PRSV-W) | [80] |
A. thaliana | NHEJ | eIF(iso)4E | Turnip mosaic virus (TuMV) resistance | [84] |
Rice (IR24) | NHEJ | OsSWEET13 | Bacterial blight disease resistance | [85] |
Bread wheat | NHEJ | TaMLO-A1, TaMLO-B1, and TaMLOD1 | Powdery mildew resistance | [86] |
Maize | HDR | ARGOS8 | Increased grain yield under drought stress | [87] |
Tomato | NHEJ | SlMAPK3 | Drought tolerance | [88] |
A. thaliana | HDR | MIR169a | Drought tolerance | [89] |
A. thaliana | NHEJ | OST2 (OPEN STOMATA 2) (AHA1) | Increased stomatal closure in response to abscisic acid (ABA), | [90] |
Rice | HDR/ NHEJ | OsPDS, OsMPK2, OsBADH2 | Involved in various abiotic stress tolerance | [91] |
Rice | NHEJ | OsMPK5 | Various abiotic stress tolerance and disease resistance | [92] |
Rice | NHEJ/ HDR | OsMPK2, OsDEP1 | Yield under stress | [93] |
Crop | Method | Target Gene | Stress/Trait | References |
---|---|---|---|---|
Rice | NHEJ | 2.5604 gRNA for 12,802 genes | Creating genome wide mutant library | [95] |
Maize | NHEJ | ZmIPK1A ZmIPK and ZmMRP4 | Phytic acid synthesis | [96] |
Wheat | HDR | TaVIT2 | Fe content | [97] |
Soybean | NHEJ | GmPDS11 and GmPDS18 | Carotenoid biosynthesis | [98] |
Tomato | NHEJ | Rin | Fruit ripening | [99] |
Potato | HDR | ALS1 | Herbicide resistance | [100] |
Cassava | NHEJ | MePDS | Carotenoid biosynthesis | [101] |
Rice | NHEJ | 2.5604 gRNA for 12,802 genes | Creating genome wide mutant library | [95] |
Maize | NHEJ | ZmIPK1A ZmIPK and ZmMRP4 | Phytic acid synthesis | [96] |
Wheat | HDR | TaVIT2 | Fe content | [97] |
Soybean | NHEJ | GmPDS11 and GmPDS18 | Carotenoid biosynthesis | [98] |
Tomato | NHEJ | Rin | Fruit ripening | [99] |
Potato | HDR | ALS1 | Herbicide resistance | [100] |
Cassava | NHEJ | MePDS | Carotenoid biosynthesis | [101] |
Species | Targeted Genes | Strategy | Germline Transmission Rate (%) | G1 Mutation Rate (%) | References |
---|---|---|---|---|---|
Drosophila spec. | yellow, whit | mRNA INJ | 0–79 | 0–34.5 | [116] |
CG4221, CG5961, Chameau | mRNA INJ with donor | 8.1–26.7 | 2.7–10.4 | [117] | |
yellow | DNA INJ with donor | 5.9–20.7 | 0.25–1.37 | [115] | |
yellow | Rapid INJ with donor | 8–53 | 15 | [118] | |
Bombyx mori | BmBLOS2 | mRNA INJ | 95.5 | 35.6 | [119] |
th, re, fl, yellow-e, kynu, ebony | DNA INJ | 5.7–18.9 | ND | [120] | |
Aedes aegypti | ECFP | mRNA INJ + DNA INJ | 0 | 5.5 | [121] |
Daphnia magna | eyeless | mRNA INJ | 18–47 | 8.2 | [122] |
Tribolium castaneum | eGFP1 | mRNA INJ + DNA INJ with donor | 55–80 | 71–100 | [123] |
Papilio xuthus, P. machaon | abdominal-B, ebony, frizzled | mRNA INJ | 18.33–90.85 | ND | [124] |
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Tavakoli, K.; Pour-Aboughadareh, A.; Kianersi, F.; Poczai, P.; Etminan, A.; Shooshtari, L. Applications of CRISPR-Cas9 as an Advanced Genome Editing System in Life Sciences. BioTech 2021, 10, 14. https://doi.org/10.3390/biotech10030014
Tavakoli K, Pour-Aboughadareh A, Kianersi F, Poczai P, Etminan A, Shooshtari L. Applications of CRISPR-Cas9 as an Advanced Genome Editing System in Life Sciences. BioTech. 2021; 10(3):14. https://doi.org/10.3390/biotech10030014
Chicago/Turabian StyleTavakoli, Kamand, Alireza Pour-Aboughadareh, Farzad Kianersi, Peter Poczai, Alireza Etminan, and Lia Shooshtari. 2021. "Applications of CRISPR-Cas9 as an Advanced Genome Editing System in Life Sciences" BioTech 10, no. 3: 14. https://doi.org/10.3390/biotech10030014