{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,28]],"date-time":"2026-04-28T12:25:26Z","timestamp":1777379126117,"version":"3.51.4"},"reference-count":74,"publisher":"SAGE Publications","issue":"1","license":[{"start":{"date-parts":[[2018,1,1]],"date-time":"2018-01-01T00:00:00Z","timestamp":1514764800000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.sagepub.com\/page\/policies\/text-and-data-mining-license"}],"content-domain":{"domain":["journals.sagepub.com"],"crossmark-restriction":true},"short-container-title":["AIDS Research and Human Retroviruses"],"published-print":{"date-parts":[[2018,1]]},"abstract":"<jats:p>\n                    Efficiency of artificial restriction enzymes toward curing HIV has only been separately examined, using differing delivery vehicles. We compared the\n                    <jats:italic toggle=\"yes\">in vitro<\/jats:italic>\n                    transduction and target-mutagenesis efficiency of consortium plasmid and adenoviral vector delivered HIV-1\n                    <jats:italic toggle=\"yes\">pol<\/jats:italic>\n                    gene targeting zinc finger nuclease (ZFN) with CRISPR\/Cas, Custom-ZFN, CRISPR-Cas-9, and plasmids and vectors (murCTSD_pZFN, pGS-U-gRNA, pCMV-Cas-D01A, Ad5-RGD); cell lines (TZM-bl and ACH-2\/J-Lat cells); and the latency reversing agents prostratin, suberoylanilide hydroxamic acid, and phorbol myristate acetate. Cell lines were grown in either Dulbecco's modified Eagle's medium or Roswell Park Memorial Institute with the antibiotics kanamycin, zeocin, and efavirenz. Efficiency was assayed by GFP\/luciferase activity and\/or validated by yeast MEL1 reporter assay, CEL1 restriction fragment assay, and quantitative reverse transcriptase\u2013polymerase chain reaction (qRT-PCR). Ad5-RGD vectors had better transduction efficiency than murCTSD and pGS-U-gRNA\/pCMV-Cas-D01A plasmids. CRISPR\/Cas9 exhibited better target-mutagenesis efficiency relative to ZFN (delivered by either plasmid or Ad5 vector) based on gel electrophoresis of\n                    <jats:italic toggle=\"yes\">pol<\/jats:italic>\n                    gene amplicons within ACH-2 and J-Lat cells. Ad-5-RGD vectors enhanced target mutagenesis of ZFN, relative to murCTSD_pZFN plasmids, to levels of CRISPR\/Cas9 plasmids. Similar reduction of luciferase activity among TZM-bl treated with Ad5-ZFN vectors relative to CRISPR\/Cas-9 and murCTSD_pZFN plasmids was observed on challenge with HIV-1. qRT-PCR of HIV-1\n                    <jats:italic toggle=\"yes\">pol<\/jats:italic>\n                    gene transcripts affirmed that Ad5 (RGD) vectors enhanced target mutagenesis of ZFN. Whereas CRISPR\/Cas-9 may possess inherent superior target-mutagenesis efficiency; the efficiency of ZFN (off-target toxicity withstanding) can be enhanced by altering delivery vehicle from plasmid to Ad5 (RGD) vectors.\n                  <\/jats:p>","DOI":"10.1089\/aid.2017.0234","type":"journal-article","created":{"date-parts":[[2017,11,29]],"date-time":"2017-11-29T01:29:05Z","timestamp":1511918945000},"page":"88-102","update-policy":"https:\/\/doi.org\/10.1177\/sage-journals-update-policy","source":"Crossref","is-referenced-by-count":7,"title":["<i>In Vitro<\/i>\n                    Transduction and Target-Mutagenesis Efficiency of HIV-1\n                    <i>pol<\/i>\n                    Gene Targeting ZFN and CRISPR\/Cas9 Delivered by Various Plasmids and\/or Vectors: Toward an HIV Cure"],"prefix":"10.1177","volume":"34","author":[{"given":"Moses","family":"Okee","sequence":"first","affiliation":[{"name":"Makerere University","place":["Uganda"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alice","family":"Bayiyana","sequence":"additional","affiliation":[{"name":"Makerere University","place":["Uganda"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Carol","family":"Musubika","sequence":"additional","affiliation":[{"name":"Makerere University","place":["Uganda"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Moses L.","family":"Joloba","sequence":"additional","affiliation":[{"name":"Makerere University","place":["Uganda"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Fred","family":"Ashaba-Katabazi","sequence":"additional","affiliation":[{"name":"Makerere University","place":["Uganda"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bernard","family":"Bagaya","sequence":"additional","affiliation":[{"name":"Makerere University","place":["Uganda"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Misaki","family":"Wayengera","sequence":"additional","affiliation":[{"name":"Makerere University","place":["Uganda"]},{"name":"Makerere University","place":["Uganda"]}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"179","published-online":{"date-parts":[[2018,1,1]]},"reference":[{"key":"e_1_3_3_2_2","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMp038194"},{"key":"e_1_3_3_3_2","volume-title":"AIDS Epidemic Update","author":"UNAIDS & World Health Organization.","year":"2015","unstructured":"UNAIDS & World Health Organization.. 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