Nuclear Localization Domains for Efficient RNA Trans-Splicing
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Solution Overview
Problem
Current RNA trans-splicing techniques suffer from low efficiency due to inadequate accumulation of RNA trans-splicing molecules at the site of action within human cells, primarily because the spliceosome-mediated RNA trans-splicing requires uneven distribution of endogenous cellular enzymes throughout the cellular nucleus.
Innovation Solution
The development of compositions and methods that incorporate a nucleic acid encoding a localization domain to promote the accumulation of trans-splicing nucleic acids in the cellular nucleus, thereby enhancing the efficiency of RNA editing by increasing the localization of these molecules to the site of action.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If RNA trans-splicing molecules are administered to human cells, then therapeutic RNA editing can occur, but the efficiency is low due to inadequate accumulation at the site of action
Solution Approach 1:
The patent applies local quality by incorporating a nuclear localization signal (NLS) domain into the trans-splicing molecule design. This NLS domain specifically directs the molecule to accumulate in the nuclear compartment where the spliceosome machinery is located, rather than being uniformly distributed throughout the cell. This localized accumulation at the specific site of action (nucleus) directly resolves the contradiction by increasing the concentration of trans-splicing molecules where they are needed, thereby improving RNA trans-splicing efficiency.
2Adaptability or versatility
If endogenous cellular enzymes are used for trans-splicing, then the process occurs naturally within the cell, but the uneven distribution of these enzymes throughout the nucleus reduces trans-splicing efficiency
Solution Approach 1:
The patent uses the nuclear localization signal (NLS) as an intermediary element that mediates the interaction between the trans-splicing molecule and the nuclear compartment. The NLS acts as a bridge that directs the trans-splicing molecule to the nucleus, thereby bringing it into proximity with the endogenous cellular enzymes (spliceosome) that are unevenly distributed within the nuclear space. This intermediary mechanism resolves the contradiction by enabling effective interaction despite the uneven distribution of enzymes.
Data Source
AI summary
Disclosed are compositions comprising a nucleic acid encoding a localization domain. The localization domain may be configured to promote accumulation of the nucleic acid in the cellular nucleus as compared to a nucleic acid without the localization domain.