Development of highly efficient CRISPR-Cas13-antimicrobials against MRSA
| Project/Area Number |
23K15377
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| Research Category |
Grant-in-Aid for Early-Career Scientists
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| Allocation Type | Multi-year Fund |
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| Review Section |
Basic Section 54030:Infectious disease medicine-related
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| Research Institution | Jichi Medical University |
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Principal Investigator |
アデリーン ヨオシーンリアン 自治医科大学, 医学部, ポスト・ドクター (20956156)
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| Project Period (FY) |
2023-04-01 – 2025-03-31
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| Project Status |
Granted (Fiscal Year 2023)
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| Budget Amount *help |
¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Fiscal Year 2024: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
Fiscal Year 2023: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
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| Keywords | CRISPR-Cas13 / engineered phage / synthetic platform / antimicrobial agent / engineered bacteriophage |
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| Outline of Research at the Start |
MRSA infections need novel treatments. Developed antimicrobial capsid that kills target bacteria in a CRISPR-Cas13a sequence-specific manner has low yield and weak bactericidal effect. This study would result in high-yield engineered phage and enhanced bactericidal activity by synthetic method.
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| Outline of Annual Research Achievements |
The purpose of the research is to enhance bactericidal efficacy against MRSA by high-yield synthetic phage loaded with CRISPR-Cas13 that specifically targets drug resistance gene mecA. In this year I have successfully established the technology that reboots phages in the L-form cells of S. aureus. I have modified the phage genome to a smaller size to increase phage yield and bactericidal activity. I have constructed smaller genome-sized synthetic phages carrying CRISPR-Cas13a, CRISPR-Cas13b, CRISPR-Cas13x and CRISPR-Cas13y and compared the bactericidal activity against MRSA. I found CRISPR-Cas13a showed the strongest activity among all the CRISPR-Cas13 subtypes. I have also started on establishing the in vivo evaluation model in the MRSA infected neonatal mouse.
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| Current Status of Research Progress |
Current Status of Research Progress
1: Research has progressed more than it was originally planned.
Reason
As I have successfully achieved my objectives of 2023 research plan, I have started to establish an in vivo evaluation model for the synthetic phages in MRSA infected neonatal mouse. This experiment requires many optimization as the MRSA infection neonatal mouse model has not been reported before and the factors of phage titre and frequency of treatment needed to be taken into consideration.
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| Strategy for Future Research Activity |
Currently, the modified synthetic phage loaded with the CRISPR-Cas13a showed the strongest bactericidal activity across all the other subtypes. However, the bactericidal activity of the orthologues of the CRISPR-Cas13a is still unknown. I will compare the bactericidal efficacy of modified synthetic phages that are loaded with different CRISPR-Cas13a orthologues. I will then evaluate the efficacy of the phages in the established MRSA neonatal mouse. As a result from this plan, I hope to successfully develop a highly efficient CRISPR-Cas13-antimicrobials agent against MRSA that can be used in the clinical setting.
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Report
(1 results)
Research Products
(1 results)
