☆ 4.7 Article

Generation of T cell receptor-retrogenic mice: improved retroviral-mediated stem cell gene transfer

NATURE PROTOCOLS (2013)

期刊

NATURE PROTOCOLS

卷 8, 期 10, 页码 1837-1840

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NATURE PUBLISHING GROUP

DOI: 10.1038/nprot.2013.111

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Biochemical Research Methods

资金

NIAID NIH HHS [R01 AI052199, T32 AI007610] Funding Source: Medline
NIDDK NIH HHS [R01 DK099317, R00 DK080885, R01 DK089125] Funding Source: Medline

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摘要

T he use of retrogenic mice offers a rapid and flexible approach to T cell receptor (TCRTCRTCR)-transgenic mice. By transducing bone marrow progenitor cells with a retrovirus that encodes a given TCRTCRTCR-a/b subunit, TCRTCRTCR-retrogenic mice can be generated in as few as 4-6 weeks, whereas conventional TCRTCRTCR transgenics can take 6 months or longer. In this updated protocol, we have increased the efficiency of the bone marrow transduction and bone marrow reconstitution compared with our previously published protocol. The main departure from the previous protocol is the implementation of spin transduction with the viral supernatant instead of coculture with the viral producer cell line. The changes in this protocol improve bone marrow viability, increase consistency of the bone marrow transduction and bone marrow engraftment, and they reduce the ratio of bone marrow donor mice to bone marrow recipients.

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