2023 Fiscal Year Research-status Report
Development of highly efficient CRISPR-Cas13-antimicrobials against MRSA
| Project/Area Number |
23K15377
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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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| Keywords | CRISPR-Cas13 / engineered phage / synthetic platform / antimicrobial agent |
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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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| Causes of Carryover |
The in vitro evaluation of the CRISPR-Cas13 in the year 2023 went on smoothly and was achieved faster than expected. The financial budget used was less than expected for the first year. However, in 2024, the in vivo neonatal mouse experiment requires more budget, hence the budget will be moved to 2024 to be used for the in vivo evaluation system.
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Research Products
(1 results)
