Engineered DNA Binding Proteins for Klotho Gene Expression
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
There is a need for compositions and methods to modulate Klotho activity, particularly to increase Klotho expression, given its roles in aging, cognition, oxidative stress, neurological protection, and various disorders.
Innovation Solution
Engineered DNA binding proteins are developed to target specific sequences upstream of the Klotho gene translation start site, fused with transcriptional activation domains to enhance Klotho gene expression by binding to defined target sequences within or near the Klotho gene.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional methods are used to modulate gene expression, then general applicability is maintained, but specificity to target the Klotho gene is insufficient
Solution Approach 1:
The DNA binding protein is divided into distinct functional modules: a DNA binding domain (such as zinc fingers or TALEs) that specifically recognizes the Klotho gene promoter sequence, and a transcriptional activation domain (such as VP16 or p65) that drives gene expression. This segmentation allows independent optimization of specificity and activation function.
Solution Approach 2:
The invention targets a specific local region of the Klotho gene promoter (sequences upstream of the translation start site) with high affinity binding. The DNA binding protein is engineered to recognize and bind specifically to this local target sequence, concentrating the activation effect precisely where needed to enhance Klotho expression without affecting other genes.
2Reliability
If Klotho expression is increased to provide therapeutic benefits, then protective functions are enhanced, but potential off-target effects may occur
Solution Approach 1:
The DNA binding protein is designed with a predetermined specificity for the Klotho gene promoter sequence before administration. The binding domain is engineered in advance to recognize only the specific target sequence upstream of the Klotho translation start site, preventing off-target binding and ensuring that Klotho expression enhancement occurs only at the intended locus.
Solution Approach 2:
The DNA binding protein acts as an intermediary between the transcriptional activation domain and the Klotho gene promoter. This intermediary structure ensures that activation is transmitted only to the correct target gene through specific DNA sequence recognition, while the activation domain itself remains sequestered until proper target engagement occurs.
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
The approach effectively increases Klotho expression, potentially offering therapeutic benefits for conditions such as cancer, muscle disorders, kidney disorders, cognitive enhancement, and neurological disorders by promoting Klotho's protective and regulatory functions.
Implementation Method 1
engineered DNA binding proteins that target sequences upstream of the Klotho translation start site
Implementation Method 2
By fusing the DNA binding proteins to transcriptional activations domain, the inventors successfully increased the expression of Klotho
Data Source
AI summary
The present disclosure relates to compositions and methods for modulating gene expression and in particular to DNA binding proteins and their use for increasing expression of Klotho. In some examples, the present disclosure provides an isolated or recombinant DNA binding protein comprising or attached to a transcriptional activation domain wherein the DNA binding protein binds to a target sequence within or near a Klotho gene.