| RRC ID |
37020
|
| Author |
Ebina H, Kanemura Y, Misawa N, Sakuma T, Kobayashi T, Yamamoto T, Koyanagi Y.
|
| Title |
A high excision potential of TALENs for integrated DNA of HIV-based lentiviral vector.
|
| Journal |
PLoS One
|
| Abstract |
DNA-editing technology has made it possible to rewrite genetic information in living cells. Human immunodeficiency virus (HIV) provirus, an integrated form of viral complementary DNA in host chromosomes, could be a potential target for this technology. We recently reported that HIV proviral DNA could be excised from the chromosomal DNA of HIV-based lentiviral DNA-transduced T cells after multiple introductions of a clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9 endonuclease system targeting HIV long terminal repeats (LTR). Here, we generated a more efficient strategy that enables the excision of HIV proviral DNA using customized transcription activator-like effector nucleases (TALENs) targeting the same HIV LTR site. A single transfection of TALEN-encoding mRNA, prepared from in vitro transcription, resulted in more than 80% of lentiviral vector DNA being successfully removed from the T cell lines. Furthermore, we developed a lentiviral vector system that takes advantage of the efficient proviral excision with TALENs and permits the simple selection of gene-transduced and excised cells in T cell lines.
|
| Volume |
10(3)
|
| Pages |
e0120047
|
| Published |
2015-1-1
|
| DOI |
10.1371/journal.pone.0120047
|
| PII |
PONE-D-14-49017
|
| PMID |
25781496
|
| PMC |
PMC4363575
|
| MeSH |
CRISPR-Cas Systems / genetics*
Cell Line
DNA, Viral / genetics*
DNA, Viral / metabolism
Genetic Vectors / genetics*
Genetic Vectors / metabolism
HIV-1 / genetics*
HIV-1 / metabolism
Humans
Transduction, Genetic*
Virus Integration / genetics*
|
| IF |
2.74
|
| Times Cited |
39
|
|
WOS Category
|
BIOTECHNOLOGY & APPLIED MICROBIOLOGY
|
| Resource |
| Human and Animal Cells |
293T(RCB2202) |
