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Journal Article

Group II Introns Designed to Insert into Therapeutically Relevant DNA Target Sites in Human Cells

Huatao Guo, Michael Karberg, Meredith Long, J. P. Jones III, Bruce Sullenger and Alan M. Lambowitz
Science
New Series, Vol. 289, No. 5478 (Jul. 21, 2000), pp. 452-457
Stable URL: http://www.jstor.org/stable/3077378
Page Count: 6

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Topics: Introns, DNA, Plasmids, RNA, HIV 1, Exons, Nucleotides, Polymerase chain reaction, Genetics, T lymphocytes
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Group II Introns Designed to Insert into Therapeutically Relevant DNA Target Sites in Human Cells
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Abstract

Mobile group II intron RNAs insert directly into DNA target sites and are then reverse-transcribed into genomic DNA by the associated intron-encoded protein. Target site recognition involves modifiable base-pairing interactions between the intron RNA and a >14-nucleotide region of the DNA target site, as well as fixed interactions between the protein and flanking regions. Here, we developed a highly efficient Escherichia coli genetic assay to determine detailed target site recognition rules for the Lactococcus lactis group II intron Ll.LtrB and to select introns that insert into desired target sites. Using human immunodeficiency virus-type 1 (HIV-1) proviral DNA and the human CCR5 gene as examples, we show that group II introns can be retargeted to insert efficiently into virtually any target DNA and that the retargeted introns retain activity in human cells. This work provides the practical basis for potential applications of targeted group II introns in genetic engineering, functional genomics, and gene therapy.

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