Adding Inosine at level 3 ?

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Inosine is a modified adenine. It can bond with C U and A (i dont know associated Free energy)
My idea is to add this base when players reach 30000points for exemple.
What do you think about it?
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yannma

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  • excited

Posted 7 years ago

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rhiju, Researcher

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putting in inosine and other 'non-standard' bases would be very neat, but we can only introduce those into RNA if we chemically synthesize it which is very expensive and difficult for long RNAs.

We currently use an enzyme to transcribe the RNA from DNA, which is cheap, but keeps us limited to the usual four nucleotides...
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yannma

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Thanks for your anwer,

I know its not realy easy, but to make it cheaper than manking it ful chemicaly, isnt it possible to create it with an "hybrid technik" ...

For exemple approximately ,
to create 3'(1)-AAUUGGIGGUUAA-(2)5'
-Is it possible to synthetise from DNA :
(1)-AAUUGG and GGUUAA(2), (two part of wanted RNA without inosine)
(-1)-UUAACCACCAAUU(-2) (a complementary of wanted RNA)
-Then chimically : Add one inosine to (1)-AAUUGG + ITP = (1)-AAUUGGI
Finaly (1)-AAUUGGI + GGUUAA(2) + (-1)-UUAACCACCAAUU(-2) + Ligase

Is this impossible ? Wrong ? Not so cheap (Ligase isnt an expensive enzyme isnt it ?) ? )
Is nt it possible to add chimically one isolated base (and only one) next to an RNA ?

Thanks again for your attention.
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yannma

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perhaps should i learn how you create your RNA in detail before thinking about this ^^'
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rhiju, Researcher

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Yes, you are completely correct -- instead of chemical synthesis, we could enzymatically synthesize pieces separately, add on I (or have some regions where all G's are substituted for I's), and then ligate, and then purify. Your procedure would work! And indeed we and collaborators have carried this out for other 'special' bases like fluorescent analogs.

The primary issue is the time and care involved -- and the fact that the protocol would be different for any arbitrary sequence. That means that we couldn't include it as part of our 'massively parallel' workflow which enables us to dozens (and soon thousands) of synthesize arbitrary player sequences. That being said, we'll keep it in mind -- there might be a way to put inosine or other bases on the 3' end or the 5' end of arbitrary sequences as part of the workflow, and we hadn't considered that before your post.