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Comparative Analysis of piggyBac, CRISPR/Cas9 and TALEN Mediated BAC Transgenesis in the Zygote for the Generation of Humanized SIRPA Rats
BAC transgenic mammalian systems offer an important platform for recapitulating human gene expression and disease modeling. While the larger body mass, and greater genetic and physiologic similarity to humans render rats well suited for reproducing human immune diseases and evaluating therapeutic st...
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Published in: | Scientific reports 2016-08, Vol.6 (1), p.31455-31455, Article 31455 |
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Main Authors: | , , , , , , , , , , , |
Format: | Article |
Language: | English |
Subjects: | |
Citations: | Items that this one cites Items that cite this one |
Online Access: | Get full text |
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Summary: | BAC transgenic mammalian systems offer an important platform for recapitulating human gene expression and disease modeling. While the larger body mass, and greater genetic and physiologic similarity to humans render rats well suited for reproducing human immune diseases and evaluating therapeutic strategies, difficulties of generating BAC transgenic rats have hindered progress. Thus, an efficient method for BAC transgenesis in rats would be valuable. Immunodeficient mice carrying a human
SIRPA
transgene have previously been shown to support improved human cell hematopoiesis. Here, we have generated for the first time, human
SIRPA
BAC transgenic rats, for which the gene is faithfully expressed, functionally active, and germline transmissible. To do this, human
SIRPA
BAC was modified with elements to work in coordination with genome engineering technologies-
piggyBac
, CRISPR/Cas9 or TALEN. Our findings show that
piggyBac
transposition is a more efficient approach than the classical BAC transgenesis, resulting in complete BAC integration with predictable end sequences, thereby permitting precise assessment of the integration site. Neither CRISPR/Cas9 nor TALEN increased BAC transgenesis. Therefore, an efficient generation of human
SIRPA
transgenic rats using
piggyBac
opens opportunities for expansion of humanized transgenic rat models in the future to advance biomedical research and therapeutic applications. |
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ISSN: | 2045-2322 2045-2322 |
DOI: | 10.1038/srep31455 |