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In vitro selection of a 3' terminal short protector that stabilizes transcripts to improve the translation efficiency in a wheat germ extract.

Abstract：

:Wheat cell-free expression systems based on wheat germ extract (WGE) enable us to briefly synthesize various types of proteins in vitro merely by exogenously adding their mRNA templates. Moreover, it is possible to produce larger amounts of protein by thoroughly removing the endosperm, which contains many translation inhibitors, including ribonucleases (RNases). However, because small amounts of RNases are also present even in an endosperm-free, high-quality WGE (hqWGE), the in-vitro transcribed mRNA is rapidly degraded. In particular, 3' exonucleases have been considered as the major RNases that degrade mRNA. We thus herein performed in vitro selection to find an effective, short 3' protector sequence from a random RNA pool. The selected sequences stabilized in vitro transcripts in the hqWGE more effectively than the previously reported, longer 3' protectors did. In addition, when one of these 3' protectors was minimized and then fused to mRNA, the translation efficiency increased 5-6-fold in the hqWGE, mainly due to the mRNA stabilization.

journal_name

Bioorg Med Chem Lett

journal_title

Bioorganic & medicinal chemistry letters

authors

Ogawa A,Kutsuna A,Takamatsu M,Okuzono T

doi

10.1016/j.bmcl.2019.06.058

subject

Has Abstract

pub_date

2019-08-15 00:00:00

pages

2141-2144

issue

16

eissn

0960-894X

issn

1464-3405

pii

S0960-894X(19)30440-8

journal_volume

29

pub_type

信件

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