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Methods for modifying translation

A technology of interaction strength, nucleic acid molecules, applied in the field of translation optimization, which can solve problems such as lack of rRNA-mRNA

Pending Publication Date: 2022-01-04
RAMOT AT TEL AVIV UNIV LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, a comprehensive understanding of rRNA-mRNA interactions is lacking, and methods to optimize mRNA sequences to enhance or reduce translation are greatly needed

Method used

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  • Methods for modifying translation
  • Methods for modifying translation
  • Methods for modifying translation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0410] Example 1: Support for selection of strong rRNA-mRNA interactions at the end of the 5'UTR and at the beginning of the coding region to regulate translation initiation and early translation elongation

[0411] First, we analyzed the 5'UTRs of 551 bacterial species with aSD (anti-Shine Delgarno) sequences in rRNA. It was shown that translation initiation in prokaryotes is initiated by the hybridization of 16S rRNA to mRNA. 16S rRNA binds to the 5'UTR near and upstream of start codon 4, such as Figure 1C shown. In fact, if Figure 1B (Black box) It can be seen that in almost all bacteria analyzed there is a significant signal supporting selection for strong rRNA-mRNA interactions at positions -8 to -17 relative to the start codon, consistent with the Shine-Dalgarno model.

[0412] A second signal supporting the selection of a strong rRNA-mRNA interaction occurs in the last nucleotide of the 5' UTR and the first five nucleotides of the coding sequence ( Figure 1B , bl...

Embodiment 2

[0415] Example 2: Selection against strong rRNA-mRNA interactions in coding regions, which prevents slowing down of translation elongation

[0416] Ribo-seq analysis in Escherichia coli revealed that strong interactions between 16S rRNA and mRNA can lead to pauses during translation elongation, hindering translation ( Figure 2D ). Therefore, avoidance of such strong rRNA-mRNA interactions in coding regions should allow ribosomes to flow efficiently during translation elongation. The deleterious effect of such strong rRNA-mRNA interacting sequences may also be due to their role in promoting internal translation initiation, which would generate truncated and frameshifted protein products. This notion is supported by the observation that the occurrence of the AUG initiation codon is significantly depleted downstream of existing strong rRNA-mRNA interaction sequences in E. coli.

[0417] Our analysis revealed evidence against a significant selection of strong rRNA-mRNA interact...

Embodiment 3

[0421] Example 3: Support for selection of strong rRNA-mRNA interactions at the ends of coding sequences to improve the fidelity of translation termination

[0422] In 82% of the analyzed bacterial species, in the 50% of positions in the last 20 nucleotides of the coding region, there was selection in favor of a strong rRNA-mRNA interaction ( Figure 3A ). This constitutes a mechanism to slow ribosome movement when approaching a stop codon and serves to ensure efficient and accurate termination and prevent translational read-through ( Figure 3F ). It is possible that this selection may have the function of assisting the initiation of overlapping or close downstream genes in operons; however, we observed this phenomenon generally across all genes and in bacteria, including operons where other genes were not immediately followed The last gene of ( Figure 3F ).

[0423] Many genes in bacteria are transcribed as operons. Specifically, in E. coli, 55% of genes are grouped in...

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Abstract

Nucleic acid molecules comprising a mutation that mutation modulates the interaction strength of the nucleic acid molecule to a 16S ribosomal RNA are provided. Methods of improving the translation process of a nucleic acid molecule and producing a nucleic acid molecule optimized for translation, as well as cells comprising the nucleic acid molecules are also provided.

Description

[0001] Cross References to Related Applications [0002] This application claims the benefit of priority to U.S. Provisional Patent Application No. 62 / 825,143, filed March 28, 2019, the contents of which are incorporated herein by reference in their entirety. technical field [0003] The present invention relates to the field of translation optimization. Background technique [0004] The region approximately 8-10 nucleotides upstream of the translation initiation site in prokaryotic mRNA tends to include purine-rich sequences. This sequence is named the Shine-Dalgarno (SD) sequence or ribosome binding site (RBS), and is believed to pass through complementary sequence bases in the 16S rRNA component of the small ribosomal subunit (i.e., the anti-Shine-Dalgarno sequence (aSD)) pairing is involved in prokaryotic translation initiation. [0005] Recent studies have also shown that sequences (motifs) within the coding region that interact with aSD, similar to SD, can slow or pa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/67C12N15/10
CPCC12N15/67C12N15/1089
Inventor T·图勒S·巴伊里B·艾普特
Owner RAMOT AT TEL AVIV UNIV LTD