Substitution of adeno-associated virus Rep protein binding and nicking sites with human Chromosome 19 sequences
<p>Abstract</p> <p>Background</p> <p>Adeno-associated virus type 2 (AAV2) preferentially integrates its DNA at a ~2 kb region of human chromosome 19, designated <it>AAVS1 </it>(also known as <it>MBS85</it>). Integration at <it>AAVS1 </it...
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| Format: | Article |
| Language: | English |
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BMC
2010-09-01
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| Series: | Virology Journal |
| Online Access: | http://www.virologyj.com/content/7/1/218 |
| _version_ | 1818766624806666240 |
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| author | McAlister Victor J Owens Roland A |
| author_facet | McAlister Victor J Owens Roland A |
| author_sort | McAlister Victor J |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>Adeno-associated virus type 2 (AAV2) preferentially integrates its DNA at a ~2 kb region of human chromosome 19, designated <it>AAVS1 </it>(also known as <it>MBS85</it>). Integration at <it>AAVS1 </it>requires the AAV2 replication (Rep) proteins and a DNA sequence within <it>AAVS1 </it>containing a 16 bp Rep recognition sequence (RRS) and closely spaced Rep nicking site (also referred to as a terminal resolution site, or <it>trs</it>). The AAV2 genome is flanked by inverted terminal repeats (ITRs). Each ITR contains an RRS and closely spaced <it>trs</it>, but the sequences differ from those in <it>AAVS1</it>. These ITR sequences are required for replication and packaging.</p> <p>Results</p> <p>In this study we demonstrate that the <it>AAVS1 </it>RRS and <it>trs </it>can function in AAV2 replication, packaging and integration by replacing a 61 bp region of the AAV2 ITR with a 49 bp segment of <it>AAVS1 </it>DNA. Modifying one or both ITRs did not have a large effect on the overall virus titers. These modifications did not detectably affect integration at <it>AAVS1</it>, as measured by semi-quantitative nested PCR assays. Sequencing of integration junctions shows the joining of the modified ITRs to <it>AAVS1 </it>sequences.</p> <p>Conclusions</p> <p>The ability of these <it>AAVS1 </it>sequences to substitute for the AAV2 RRS and <it>trs </it>provides indirect evidence that the stable secondary structure encompassing the <it>trs </it>is part of the AAV2 packaging signal.</p> |
| first_indexed | 2024-12-18T08:36:57Z |
| format | Article |
| id | doaj.art-86806d9f54064169b63b3781b0e3dd19 |
| institution | Directory Open Access Journal |
| issn | 1743-422X |
| language | English |
| last_indexed | 2024-12-18T08:36:57Z |
| publishDate | 2010-09-01 |
| publisher | BMC |
| record_format | Article |
| series | Virology Journal |
| spelling | doaj.art-86806d9f54064169b63b3781b0e3dd192022-12-21T21:14:18ZengBMCVirology Journal1743-422X2010-09-017121810.1186/1743-422X-7-218Substitution of adeno-associated virus Rep protein binding and nicking sites with human Chromosome 19 sequencesMcAlister Victor JOwens Roland A<p>Abstract</p> <p>Background</p> <p>Adeno-associated virus type 2 (AAV2) preferentially integrates its DNA at a ~2 kb region of human chromosome 19, designated <it>AAVS1 </it>(also known as <it>MBS85</it>). Integration at <it>AAVS1 </it>requires the AAV2 replication (Rep) proteins and a DNA sequence within <it>AAVS1 </it>containing a 16 bp Rep recognition sequence (RRS) and closely spaced Rep nicking site (also referred to as a terminal resolution site, or <it>trs</it>). The AAV2 genome is flanked by inverted terminal repeats (ITRs). Each ITR contains an RRS and closely spaced <it>trs</it>, but the sequences differ from those in <it>AAVS1</it>. These ITR sequences are required for replication and packaging.</p> <p>Results</p> <p>In this study we demonstrate that the <it>AAVS1 </it>RRS and <it>trs </it>can function in AAV2 replication, packaging and integration by replacing a 61 bp region of the AAV2 ITR with a 49 bp segment of <it>AAVS1 </it>DNA. Modifying one or both ITRs did not have a large effect on the overall virus titers. These modifications did not detectably affect integration at <it>AAVS1</it>, as measured by semi-quantitative nested PCR assays. Sequencing of integration junctions shows the joining of the modified ITRs to <it>AAVS1 </it>sequences.</p> <p>Conclusions</p> <p>The ability of these <it>AAVS1 </it>sequences to substitute for the AAV2 RRS and <it>trs </it>provides indirect evidence that the stable secondary structure encompassing the <it>trs </it>is part of the AAV2 packaging signal.</p>http://www.virologyj.com/content/7/1/218 |
| spellingShingle | McAlister Victor J Owens Roland A Substitution of adeno-associated virus Rep protein binding and nicking sites with human Chromosome 19 sequences Virology Journal |
| title | Substitution of adeno-associated virus Rep protein binding and nicking sites with human Chromosome 19 sequences |
| title_full | Substitution of adeno-associated virus Rep protein binding and nicking sites with human Chromosome 19 sequences |
| title_fullStr | Substitution of adeno-associated virus Rep protein binding and nicking sites with human Chromosome 19 sequences |
| title_full_unstemmed | Substitution of adeno-associated virus Rep protein binding and nicking sites with human Chromosome 19 sequences |
| title_short | Substitution of adeno-associated virus Rep protein binding and nicking sites with human Chromosome 19 sequences |
| title_sort | substitution of adeno associated virus rep protein binding and nicking sites with human chromosome 19 sequences |
| url | http://www.virologyj.com/content/7/1/218 |
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