IRES2 vs 2A peptide

Dario

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Dec 23, 2016, 6:24:11 AM12/23/16

to Genome Engineering using CRISPR/Cas Systems

Hi Everyone,

a couple of quick question. Does anyone have any data on how less efficient is IRES2 compare to a 2A peptide for a reporter gene?
Second question would be, where is the best place to put an IRES2 sequence in an endogenous locus? right after 3'UTR?

Thanks for helping.

Best and Merry Christmas.

Dario

Jonathan Geisinger

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Jan 3, 2017, 7:52:36 PM1/3/17

to Genome Engineering using CRISPR/Cas Systems

In my experience, IRES is worse by 4-5-fold compared to 2A skipping peptides (specifically P2A).

saul kivimae

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Jan 4, 2017, 6:25:19 PM1/4/17

to Genome Engineering using CRISPR/Cas Systems

2A always has some fusion protein. If that matters then IRES2 is a better choice. How bad is the expression of the downstream ORF is very case dependent. I have had good success with IRES2. If you are limited by sequence length in a virus then 2A saves space though.

Cheers
Saul

Dario

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Jan 5, 2017, 8:01:36 AM1/5/17

to Genome Engineering using CRISPR/Cas Systems

Thanks guys for your inputs.

Cheers,
Dario

mario

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Jan 7, 2017, 2:58:46 PM1/7/17

to Genome Engineering using CRISPR/Cas Systems

This paper should probably answer to your question
Ciao
Mario
http://nar.oxfordjournals.org/content/early/2015/12/09/nar.gkv1338.full

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