Inserting cDNA into a retroviral vector?..

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x555
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Inserting cDNA into a retroviral vector?..

Post by x555 » Sun Mar 10, 2013 1:43 pm

Hello everyone.

Let's say I have a retroviral vector and its genome consists of single stranded RNA. Is it possible to insert double stranded cDNA into this vector? If yes, what are the processes that need to happen in order to make this occur? And can I use DNA ligase to seal the fragments of cDNA and retroviral vector?

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JackBean
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Post by JackBean » Mon Mar 11, 2013 12:20 am

you will hardly insert dsDNA into ssRNA. You have to make RNA from it and that use for the insertion. But you have to be aware whether your retrovirus contains + or - RNA strand.
http://www.biolib.cz/en/main/

Cis or trans? That's what matters.

x555
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Post by x555 » Mon Mar 11, 2013 7:59 am

Thank you, but maybe there's another option: could I make the cDNA of the retrovirus using RT-PCR, then insert the cDNA of the gene into the vector, and later insert that vector into dividing cells of another organism? Theoretically, it's the retroviral cDNA that inserts itself into the host chromosomes, so maybe this would work?

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JackBean
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Post by JackBean » Mon Mar 11, 2013 8:50 am

sure that it done on regular basis. ut depends what cells you want to insert it into, what vector to use, etc.
http://www.biolib.cz/en/main/

Cis or trans? That's what matters.

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