Somatic Cell Nuclear Transfer in Non-Human Primates
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Solution Overview
Problem
Current methods for constructing non-human primate transgenic animal models, particularly using somatic cell nuclear transfer, face challenges such as high technical difficulty, low efficiency, and inefficiencies in achieving genetic modifications like conditional knockouts or knock-ins, leading to unsuitable first-generation models and genetic background inconsistencies.
Innovation Solution
A method involving the use of specific reprogramming activators like Kdm4d protein or its encoding mRNA, combined with activation treatment agents like ionomycin and histone deacetylase inhibitors, to optimize the somatic cell nuclear transfer process, improving the efficiency of reprogramming and generating healthy cloned non-human primate animals.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Stability of the object's composition
If somatic cell nuclear transfer is used to prepare non-human primate cloned animals, then genetic background consistency can be achieved, but the technical difficulty and operational complexity increase significantly
Solution Approach 1:
The patent applies parameter changes by optimizing activation treatment conditions (using ionomycin and 6-DMAP at specific concentrations and time points) and reprogramming conditions (TSA treatment) to improve the efficiency of somatic cell nuclear transfer in non-human primates, thereby reducing technical difficulty while maintaining genetic consistency
Solution Approach 2:
The patent employs preliminary action by pre-treating somatic cells with activation agents before nuclear transfer and using reprogramming activators during the early embryonic stages to facilitate successful cloning, which simplifies the overall technical process
2Ease of operation
If conventional activation treatment is used without reprogramming activators, then the operation process is simpler, but the development efficiency of cloned embryos is very low
Solution Approach 1:
The patent introduces reprogramming activators (such as Kdm4d protein or its encoding nucleic acids) as intermediaries to bridge the gap between simple activation treatment and efficient embryo development, enabling significant improvement in cloned embryo development efficiency without substantially complicating the operational process
Solution Approach 2:
The patent changes the biochemical parameters by introducing reprogramming activators that modify chromatin structure and gene expression patterns, thereby dramatically improving embryo development efficiency while maintaining operational simplicity
3Manufacturing precision
If lentivirus infection or editable nuclease technology is used to obtain transgenic animals, then genetic modification can be achieved, but chimerism occurs in F0 generation making them unsuitable for research
Solution Approach 1:
The patent applies preliminary action by performing genetic modifications in somatic cells before nuclear transfer, ensuring that the resulting cloned animals are non-chimeric F0 generation models suitable for research, thereby resolving the reliability issue while maintaining genetic modification precision
Solution Approach 2:
The patent extracts the genetic modification step from the embryo manipulation process and performs it separately in somatic cells, which eliminates chimerism and produces reliable non-human primate models suitable for scientific research
4Manufacturing precision
If editable nucleases are injected into embryos for complex genetic modifications, then gene knock-in or conditional knockout can be achieved, but the efficiency is very low at mammalian level
Solution Approach 1:
The patent performs complex genetic modifications in somatic cells before nuclear transfer, which significantly improves the efficiency of gene knock-in and conditional knockout compared to direct embryo injection, while maintaining the precision of complex genetic modifications
Solution Approach 2:
The patent uses somatic cells as an intermediary system to perform complex genetic modifications with high efficiency, then transfers the modified nucleus to enucleated oocytes, achieving both high productivity and precision in complex genetic modifications
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 significantly enhances the success rate of somatic cell nuclear transfer in non-human primates, allowing for the production of viable cloned animals and overcoming previous inefficiencies, enabling consistent genetic backgrounds and reducing chimerism.
Implementation Method 1
activation treatment agents like ionomycin
Implementation Method 2
histone deacetylase inhibitors
Implementation Method 3
specific reprogramming activators like Kdm4d protein
Data Source
AI summary
Provided for the first time in the present invention is a method for preparing a non-human primate somatic cell cloned animal, which method specifically comprises the steps of: (i) providing a reconstructed egg, wherein the egg comes from the non-human primate (ii) activating the reconstructed egg to form an activated reconstructed egg or activated reconstructed embryo formed by the reconstructed egg; (iii) reprogramming (a) the activated reconstructed egg or (b) embryonic cells of the activated reconstructed embryo to obtain a reprogrammed reconstructed egg or reprogrammed reconstructed embryo; and (iv) regenerating the reprogrammed reconstructed egg or reprogrammed reconstructed embryo to obtain the non-human primate somatic cell cloned animal. The method of the present invention can significantly improve the developmental capacity of nucleus-transplanted embryos in non-human primates (such as monkeys).