Goals/Aim:

Goals/Aim:

Lassa fever virus have infected more than thousand people with more than thousand death each year in West Africa. The fatality of Lassa fever in infected patient is more than 80%. In this project, we develop a replication-incompetent recombinant adenovirus serotype 26 housing a codon-optimized gene that contained the glycoprotein of Lassa fever virus (Ad26-GP-LFV).

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This Ad26-GP-LFV will be able to elicit antibodies production against Lassa fever virus. This is a process that has been successful in other vaccine production, like Ebola, SARs etc. The glycoprotein is the part on the Lassa fever virus that provide attachment of the virus to it host. If this Ad26-GP-LFV is successfully engineer, it can be use in future vaccine production that will save thousand life from a virus with high fatality rate.

General Plans of Action:

To construct the adenovirus vector, first I will need to prepare a replication-incompetent adenovirus by deleting E1/E3 gene from serotype 26 within day 2. The glycoprotein from Lassa fever virus will then be inserted at the E1 site of Ad26 along with human CMV promoter within day 4-6. The adenovirus-mediated GP was propagated in Human embryonic kidney (HEK) 293 cells for replication for 2-3 days.

Assessment Plans:

 

adenovirus-mediated glycoprotein of Lassa fever virus (Ad26-GP-LFV) yield was analyzed by western blot analysis. PCR performed to confirm the identity of the purified vector. DNA sequencing was used to check the complete transgene region and the flanking sequences.

 

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