RRC ID 54577
Author Yamaguchi T, Morikawa A, Miyoshi H.
Title Comparison of gene-trapping efficiency between retroviral and lentiviral vectors in mouse embryonic stem cells.
Journal Biochem. Biophys. Res. Commun.
Abstract Gene trapping is a method of inserting DNA into the genome at random, generating insertional mutations throughout the genome. The efficiency of retroviral gene trapping is not sufficient in part because of a strong preference for retroviral integration near transcription start sites. In contrast, lentiviral vectors strongly favor integration in the entire region of highly active genes, suggesting that lentiviral vectors would improve the efficiency of gene trapping. In this study, we constructed both lentiviral and retroviral gene-trap vectors and analyzed integration sites in mouse embryonic stem (ES) cells. The frequency of false-positive gene-trap events was about 12-fold higher for the retroviral vector compared to the lentiviral vector. Within intragenic regions, most of the retroviral vector integration sites were found in the 5' untranslated region, while the lentiviral vector integrated uniformly throughout transcriptional units. The trapping efficiency of unique genes was significantly higher for the lentiviral vector (~83%) than for the retroviral vector (~51%). Our data demonstrate that the lentiviral vector can trap the active genes more efficiently than the retroviral vector and will facilitate efficient generation of gene-trap libraries not only in ES cells but also in a wide variety of cell lines and primary cells.
Volume 425(2)
Pages 297-303
Published 2012-8-24
DOI 10.1016/j.bbrc.2012.07.085
PII S0006-291X(12)01392-7
PMID 22842569
MeSH Animals DNA, Intergenic / genetics Embryonic Stem Cells* Gene Expression Gene Transfer Techniques* Genetic Vectors* INDEL Mutation Lentivirus* Mice Retroviridae* Transcription Initiation Site Virus Integration*
IF 2.559
Resource
DNA material CSII-EF-MCS (RDB04378) CS-CDF-CG-PRE (RDB04379)