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INDONESIA
Molecular and Cellular Biomedical Sciences (MCBS)
ISSN : 25274384     EISSN : 25273442     DOI : -
Core Subject : Health, Science,
Molecular and Cellular Biomedical Sciences (MCBS) has been published by Cell and BioPharmaceutical Institute (CBPI), a biannually published scientific journal, is an open access, peer-reviewed journal that supports all topics in Biology, Pathology, Pharmacology, Biochemistry, Histology and Biomedicine in the aspect of molecular and cellular.
Arjuna Subject : -
Articles 132 Documents
CRISPR Target-based Single-guide RNA (sgRNA) for Diagnostic Testing of Hepatitis B Virus Jeanne Elvia Christian; Hartiyowidi Yuliawuri; Edvan Arifsaputra Suherman
Molecular and Cellular Biomedical Sciences Vol 7, No 2 (2023)
Publisher : Cell and BioPharmaceutical Institute

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21705/mcbs.v7i2.301

Abstract

Background: Indonesia is the second-highest country with hepatitis B cases in the South East Asian region. Clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR associated protein 12 (Cas12) could be developed as a diagnostic tool to detect hepatitis B infection. This study was aimed to develop a diagnostic method for hepatitis B virus by designing CRISPR target-based single-guide RNA (sgRNA).Materials and method: The preCore/Core-gene sequences of hepatitis B virus were collected from the National Center for Biotechnology Information (NCBI) website. The selected sequence was submitted to Cas Designer and CRISPOR tools to design sgRNA. The resulting sgRNA was cloned in silico into an expression vector using Benchling software.Results: The 23-nucleotide sequence 5'- GTAGTCAGTTATGTCAATGTTAA-3’ had 30% GC content, 68.3 out-of-frame and 76 predicted efficiencies. This sequence had no mismatch based on analysis.Conclusion: This preliminary study will help design a CRISPR-based diagnostic kit for the detection of hepatitis B virus in Indonesia. However, further in vitro and in vivo studies are required to demonstrate its potential and efficiency.Keywords: CRISPR-Cas12b, diagnostic, HBV, sgRNA 
The Construction of A Multi-epitope Vaccine Against Klebsiella pneumoniae Using in silico Approach Dhammiko Wonggo; Mariana Wahjudi
Molecular and Cellular Biomedical Sciences Vol 7, No 2 (2023)
Publisher : Cell and BioPharmaceutical Institute

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21705/mcbs.v7i2.343

Abstract

Background: Klebsiella pneumoniae is one of the bacteria that causes pneumonia infection. Even though the number of pneumonia cases is relatively high and has become a global problem, there is still no vaccine available to prevent this disease. This study was aimed to design a multi-epitope vaccine design through an in silico approach, against K. pneumoniae.Materials and method: Vaccine candidate was constructed based on proteins of K. pneumoniae. These proteins were analyzed to identify the antigens sequence for multi-epitope vaccine design. The constructed vaccine was predicted for allergenicity, toxicity, population coverage, and its physicochemical properties. The vaccine structure was then docked with the toll like receptor 2 (TLR2) molecule to show the interaction. Expression analysis and cloning of the constructed vaccine was carried out in the pET-28a vector using SnapGene.Results: The vaccine was 567 amino acids long, consisting of Cholera Toxin Subunit B as an adjuvant, 6 B-cell epitopes, 11 cytotoxic T-cell epitopes, and 10 helper T-cell epitopes connected with the appropriate linker. Epitopes analysis showed that the vaccine will be a non-toxic, has high antigenicity, but non-allergenic. The vaccine was predicted to be stable, hydrophilic, and had a low risk of triggering autoimmune response. The vaccine molecule was compatible to humans TLR2 molecule. Furthermore, visualization of the candidate vaccine protein on pET-28a showed that the vaccine protein might be expressed correctly.Conclusion: The construction of multi-epitope vaccine has been developed, which might be a good vaccine candidate, containing 6 B-cell epitopes, 11 CTL epitopes, and 10 HTL epitopes. The construct may help scientists to experimentally formulate multi-epitope vaccine against K. pneumoniae in the future.Keywords: in silico, Klebsiella pneumoniae, multi-epitope, vaccine