Prime Editing Complex Corrects SLC37A4 Mutations
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Solution Overview
Problem
Glycogen Storage Disease Type 1B is caused by mutations in the SLC37A4 gene, leading to impaired glucose homeostasis due to the liver's inability to break down glycogen, and current treatments are inadequate.
Innovation Solution
The use of prime editing technology to specifically target and correct mutations in the SLC37A4 gene, such as the G339C and L348fs mutations, by introducing precise nucleotide edits using prime editing guide RNAs (PEgRNAs) and a prime editor complex.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If prime editing is used to correct SLC37A4 mutations, then the accuracy of gene editing is improved, but the complexity of the editing system increases
Solution Approach 1:
The prime editing system is divided into separate functional components: the PEgRNA molecule is segmented into distinct functional regions (spacer sequence for target recognition, primer binding site for anchoring, and editing template for providing correction information). This segmentation allows each component to perform its specific function independently, improving overall editing accuracy while maintaining manageable system complexity through modular design.
Solution Approach 2:
The PEgRNA acts as an intermediary carrier that bridges the prime editor enzyme and the target DNA sequence. It mediates the interaction by providing both target recognition through its spacer sequence and the necessary template information for accurate editing through its editing template region, thereby achieving high precision without requiring direct complex enzyme-DNA interactions.
2Measurement precision
If prime editing guide RNAs are designed to target specific mutations, then the specificity of editing is improved, but the difficulty of detecting and measuring editing efficiency increases
Solution Approach 1:
The patent employs sequence-specific PEgRNAs that function as molecular probes, where the design of the guide RNA sequence itself serves as the detection mechanism. By designing PEgRNAs with unique spacer sequences complementary to specific mutant alleles, the system achieves both editing specificity and detectability through sequence analysis of the edited DNA, eliminating the need for separate detection assays.
3Adaptability or versatility
If multiple PEgRNAs are used to address different mutations, then the versatility of treatment is improved, but the complexity of the treatment protocol increases
Solution Approach 1:
The patent describes a universal prime editing platform where a single prime editor enzyme can work with multiple different PEgRNA molecules. Each PEgRNA is designed with a specific spacer sequence targeting different SLC37A4 mutations, allowing the same core editing system to address multiple disease-causing variants. This multi-functionality approach enables treatment versatility across different mutations while maintaining protocol simplicity through the use of a single editor platform.
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
This approach allows for the restoration of the wild-type sequence in the SLC37A4 gene, potentially alleviating the symptoms of Glycogen Storage Disease Type 1B by improving glucose homeostasis.
Implementation Method 1
the spacer sequence of a PEgRNA recognizes and anneals with a search target sequence in a target strand of the target gene
Implementation Method 2
A prime editing complex may generate a nick in the target gene on the edit strand
Implementation Method 3
a single stranded DNA is synthesized using an editing template of the PEgRNA as a template
Implementation Method 4
Through removal of an editing target sequence on the edit strand of the target gene and DNA repair, the intended nucleotide edit(s) included in the newly synthesized single stranded DNA are incorporated into the target SLC37A4 gene
Data Source
AI summary
Provided herein are compositions and methods of using prime editing systems comprising prime editors and prime editing guide RNAs for treatment of genetic disorders.