Safety and Efficacy of CRISPR/Cas9 mRNA Instantaneous Gene Editing Therapy to Treat Refractory Viral Keratitis

Inclusion Criteria:

Patients (replase) with refractory keratitis caused by herpes virus type I who has had at least one time failed corneal transplant.

1. Age between 18 to 70 years.
2. No systemic immune eye disease.
3. Good eyelid structure and blink function.
4. Exists the potential of visual recovery by evaluation of ocular structure and function.
5. Patients with refractory keratitis who are repeatedly infected with HSV-1 virus (more than three times per year) and resistant to topical or systemic anti-viral agents, with no response to regular immunosuppressive agents.
6. Patients who are obviously suffering from relapse HSV infections with corneal perforation, requiring corneal transplantation;
7. No history of corneal trauma.
8. Subjects or their legal guardians voluntarily participate in this study, sign informed consent, good compliance and cooperation with follow-up visits.

Exclusion Criteria:

1. Lacrimal coating and blink function loss.
2. Schirmer's test result is less than 2mm for severe dry eye disease.
3. Pregnant and lactating women (pregnancy defined in this study as positive urine pregnancy test).
4. Currently is involved in clinical trials of other drugs or medical devices.
5. Active eye infection (including but not limited to: blepharitis, infectious conjunctivitis, keratitis, sclerotitis, endophthalmitis) in target eye or contralateral eye within 30 days prior to enrollment.
6. Ocular surface malignant tumor.
7. A history of allergic reaction or allergy to sodium luciferin, allergy to protein products used for treatment or diagnosis, allergy to ≥ 2 drugs or non-drug factors, or current allergic disease.
8. current in an infectious disease requiring oral, intramuscular or intravenous administration.
9. Patients with systemic immune diseases.
10. Any uncontrolled clinical problems (such as severe mental, neurological, cardiovascular, respiratory and other systemic diseases and malignant neoplasms).
11. Not effective contraception.
12. In uncontrolled hypertension, systolic is no less than 160 mmhg, diastolic is no less than 100 mmhg.
13. In uncontrolled diabetes, fasting glucose is no less than 10.0umol/L.
14. Renal insufficiency, serum creatinine is more than 133umol/L.
15. Arrhythmia, myocardial ischemia, myocardial infarction (diagnosed by electrocardiogram).
16. Liver dysfunction, al ANINE aminotransferase and aspartate aminotransferase levels are higher than 80 IU/L.
17. Platelet level is below 100,000 /uL or above 450,000 /uL.
18. Hemoglobin level is below 10.0g/dL (male) or 9.0g/dL (female).
19. No anticoagulant was used, prothrombin time is higher than 16s, and thrombin time of activated part is higher than 50s.
20. HIV infection (HIV-positive).
21. Subjects lack compliance with the study or the ability to sign informed consent.
22. There are currently signs of systemic infection, including fever and ongoing antibiotic treatment (in this study, systemic infection was defined as deviation from normal values of white blood cells, lymphocytes, and neutrophils on routine blood tests).
23. Administration of Glucocorticoids and other systemic immunosuppressive drugs.
24. The investigator judges other conditions unsuitable for the trial