Isolated Nucleic Acid Binding Domains for Mild Nucleic Acid Release
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Solution Overview
Problem
Existing nucleic acid binding proteins (NBPs) often require harsh conditions for nucleic acid release and have limited sequence specificity, making them inefficient for broad-range binding and easy manipulation.
Innovation Solution
Isolated nucleic acid binding domains with specific amino acid sequences, capable of forming stable complexes with nucleic acids under various conditions, including temperature and ionic strength variations, without sequence specificity, and can be easily released.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If nucleic acid binding proteins are used to isolate nucleic acids, then nucleic acid binding capability is achieved, but harsh conditions (high temperature, protease digestion) are required for nucleic acid release
Solution Approach 1:
The patent extracts only the nucleic acid binding domain from the full-length nucleic acid binding protein. This isolated domain maintains the essential nucleic acid binding function while eliminating the need for harsh release conditions, allowing gentle elution at physiological temperatures and pH levels.
Solution Approach 2:
The isolated nucleic acid binding domain exhibits altered physical-chemical parameters compared to full-length proteins. It maintains stable binding across a broad range of temperatures, ionic strengths, and pH levels, enabling nucleic acid release under mild conditions without requiring extreme heat or proteolytic digestion.
2Reliability
If nucleic acid binding proteins are used, then nucleic acid isolation is achieved, but sequence specificity limits the range of applicable nucleic acids
Solution Approach 1:
The isolated nucleic acid binding domain is designed to bind nucleic acids in a non-sequence-specific manner, making it universally applicable to various types of nucleic acids including DNA, RNA, single-stranded, and double-stranded forms. This universality expands the versatility of the binding domain across different nucleic acid applications.
3Reliability
If full-length nucleic acid binding proteins are used, then stable nucleic acid binding is achieved, but the proteins form multimers that are difficult to manipulate
Solution Approach 1:
The patent segments the full-length nucleic acid binding protein into its functional binding domain, which can be isolated and expressed independently. This segmented approach produces monomeric units that are easier to manipulate and control while maintaining stable nucleic acid binding, eliminating the complexity of multimeric full-length protein structures.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
Enables efficient isolation and manipulation of nucleic acids without sequence specificity, facilitating downstream reactions and easy release under mild conditions.
Implementation Method 1
Proteins can interact with nucleic acids, such as DNA and RNA, through a variety of physical forces, such as hydrogen bonding and electrostatic interactions.
Implementation Method 2
Proteins can interact with nucleic acids, such as DNA and RNA, through a variety of physical forces, such as hydrogen bonding and electrostatic interactions.
Data Source
AI summary
Nucleic acid binding domains are described for use in isolating nucleic acid. Compositions and kits comprising these nucleic acid binding domains are also described. These nucleic acid binding domains may be used in a variety of methods.