Miskey, Csaba and Kesselring, Lisa and Querques, Irma and Abrusán, György and Barabas, Orsolya (2022) Engineered Sleeping Beauty transposase redirects transposon integration away from genes. NUCLEIC ACIDS RESEARCH, 50 (5). pp. 2807-2825. ISSN 0305-1048
|
Text
gkac092.pdf Download (6MB) | Preview |
Abstract
The Sleeping Beauty (SB) transposon system is a popular tool for genome engineering, but random integration into the genome carries a certain genotoxic risk in therapeutic applications. Here we investigate the role of amino acids H187, P247 and K248 in target site selection of the SB transposase. Structural modeling implicates these three amino acids located in positions analogous to amino acids with established functions in target site selection in retroviral integrases and transposases. Saturation mutagenesis of these residues in the SB transposase yielded variants with altered target site selection properties. Transposon integration profiling of several mutants reveals increased specificity of integrations into palindromic AT repeat target sequences in genomic regions characterized by high DNA bendability. The H187V and K248R mutants redirect integrations away from exons, transcriptional regulatory elements and nucleosomal DNA in the human genome, suggesting enhanced safety and thus utility of these SB variants in gene therapy applications.
Item Type: | Article |
---|---|
Additional Information: | Funding Agency and Grant Number: OTKAOrszagos Tudomanyos Kutatasi Alapprogramok (OTKA) [PD83571] Funding text: OTKA[PD83571 toG.A.]. Funding for open access charge: Institutional funds. |
Uncontrolled Keywords: | IMMUNODEFICIENCY-VIRUS TYPE-1; STRUCTURAL BASIS; AMINO-ACID; Elements; mutational analysis; PREFERENCES; CORE DOMAIN; TARGET SITE SELECTION; RETROVIRAL DNA INTEGRATION; |
Subjects: | Q Science / természettudomány > QH Natural history / természetrajz > QH426 Genetics / genetika, örökléstan |
SWORD Depositor: | MTMT SWORD |
Depositing User: | MTMT SWORD |
Date Deposited: | 17 Feb 2023 12:59 |
Last Modified: | 17 Feb 2023 12:59 |
URI: | http://real.mtak.hu/id/eprint/159347 |
Actions (login required)
Edit Item |