ETV2 Knockout Pig Chimeras for Humanized Blood Production
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Solution Overview
Problem
Current methods for producing human blood and vasculature rely solely on human donors, which are limited in supply and cannot be personalized to individual recipients, and there is a need for novel therapies for cardiovascular and hematopoietic diseases.
Innovation Solution
Generating ETV2 knockout pigs or other animals by deleting the ETV2 gene, then introducing human stem cells to produce human or humanized blood and vasculature, allowing for personalized tissue production and serving as a model for studying human lineage regeneration and pharmacological responses.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If human donors are used as the only source of blood and vasculature, then the supply is limited and cannot be personalized, but this is the current standard method
Solution Approach 1:
The patent uses a non-human animal (pig) as an intermediary host to produce human blood and vasculature. The pig provides the physiological environment and growth framework, while human stem cells provide the genetic identity. This intermediary system resolves the contradiction by enabling both large-scale production (pig's capacity) and human compatibility (human cell lineage).
Solution Approach 2:
The patent implements nesting by placing human stem cells inside the pig blastocyst. The human cells are nested within the non-human animal's developmental framework, allowing human blood and vasculature to be produced within a larger animal host. This enables personalized human tissues to be generated at a scale impossible through human donors alone.
2Productivity
If the Etv2 gene is deleted in pigs to create a niche for human stem cells, then human blood and vasculature can be produced, but the pig cannot produce its own blood and vasculature
Solution Approach 1:
The patent applies preliminary action by deleting the Etv2 gene in the pig before introducing human stem cells. This creates the hematopoietic niche in advance, ensuring that when human cells are introduced, they will automatically populate the blood and vasculature systems without competition from pig hematopoietic cells. The preliminary genetic modification ensures reliable human blood production.
Solution Approach 2:
The patent converts the harmful effect of Etv2 deletion (pig cannot produce its own blood) into a beneficial outcome (human cells exclusively populate the blood system). The pig's inability to produce hematopoietic cells creates a complete niche for human cells, ensuring 100% human blood and vasculature production without mixed lineage complications.
3Adaptability or versatility
If gene editing technologies are used to delete the ETV2 gene, then hematopoietic and endothelial lineages are ablated, but this requires complex genetic engineering processes
Solution Approach 1:
The patent applies the extraction principle by specifically removing the Etv2 gene from the pig genome using gene editing technologies. This targeted extraction of the critical hematopoietic regulator creates a precise niche for human stem cells. The specificity of taking out only this one gene (rather than multiple genes or using broader methods) achieves lineage ablation with relatively focused genetic engineering.
Data Source
AI summary
Described herein is a method for producing a chimeric non-human animal expressing a human ETV2 gene comprising: a) generating an ETV2 null non-human animal cell, wherein both copies of the non-human ETV2 gene carry a mutation that prevents production of functional ETV2 protein in said non-human animal; b) creating an ETV2 null non-human blastocyst by somatic cell nuclear transfer comprising fusing a nucleus from said ETV2 null non-human animal cell of a) into an enucleated non-human oocyte and activating said oocyte to divide so as to form an ETV2 null non-human blastocyst; c) introducing human stem cells into the ETV2 null non-human blastocyst of b); and d) implanting said blastocyst from c) into a pseudopregnant surrogate non-human animal to generate a chimeric non-human animal expressing human ETV2.