A Novel MutantloxP Containing Part of Long Terminal Repeat of HIV-1 in Spacer Region: Presentation of Possible Target Site for Antiviral Strategy Using Site-Specific Recombinase

Lee, Young-sam; Park, Jong‐Sang

doi:10.1006/bbrc.1998.9814

Cited by 9 publications

(5 citation statements)

References 19 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…For example, as has been noted (26), the integrated DNA copy of a retrovirus such as HIV might be excised and thereby inactivated by recombination between identical sites in the long terminal repeats.…”

Section: Discussionmentioning

confidence: 99%

Chimeric recombinases with designed DNA sequence recognition

Akopian

Boocock

et al. 2003

Proc. Natl. Acad. Sci. U.S.A.

107

View full text Add to dashboard Cite

Site-specific recombination typically occurs only between DNA sequences that have co-evolved with a natural recombinase enzyme to optimize sequence recognition, catalytic efficiency, and regulation. Here, we show that the sequence recognition and the catalysis functions of a recombinase can be specified by unrelated protein domains. We describe chimeric recombinases with a catalytic domain from an activated multiple mutant of the bacterial enzyme Tn3 resolvase, fused to a DNA recognition domain from the mouse transcription factor Zif268. These proteins catalyze efficient recombination specifically at synthetic target sites recognized by two Zif268 domains. Our results demonstrate the functional autonomy of the resolvase catalytic domain and open the way to creating ''custom-built'' recombinases that act at chosen natural target sequences.

show abstract

Section: Discussionmentioning

confidence: 99%

Chimeric recombinases with designed DNA sequence recognition

Akopian

Boocock

et al. 2003

Proc. Natl. Acad. Sci. U.S.A.

107

View full text Add to dashboard Cite

show abstract

“…Error bar indicates the standard deviation. products [Lee and Park, 1998;Lee and Saito, 1998]. This result indicates that it is not the case that strong binding af®nity is needed to operate ef®cient catalysis.…”

Section: Characterization Of Arg259 and Gln90mentioning

confidence: 99%

“…Wild-and mutant-type Cre were puri®ed essentially as described by Abremski and Hoess [Abremski and Hoess, 1984] with slight modi®cation [Lee and Park, 1998;Lee and Saito, 1998]. Wild-type Cre and most Cre mutants were eluted with 0.6 Â TSEG (X Â TSEG 20 mM Tris±HCl, pH 7.5, 1 mM EDTA, 10% (v/v) glycerol, and X is the molar concentration of NaCl [Abremski et al, 1983]) from phosphocellulose column (Whatman).…”

Section: Puri®cation Of Cre Proteinmentioning

confidence: 99%

“…The loxIL2 and loxIL3 recombination was determined by the restriction enzyme analysis [Abremski et al, 1983] with slight modi®cation [Lee and Park, 1998;Lee and Saito, 1998]. Brie¯y, recombination reactions were carried out in a 50 ml containing 50 mM Tris±HCl, pH 7.5, 10 mM MgCl 2 , 30 mM NaCl, 0.5 mg/ml bovine serum albumin, 0.5 mg puri®ed wildtype Cre, and 2 mg loxP, loxIL1, loxIL2 or loxIL3-ligated plasmid DNA (pRH43, pYLIL1, pYLIL2, or pYLIL3, respectively).…”

Section: In Vitro Recombination Assaymentioning

confidence: 99%

See 1 more Smart Citation

Characterization of Cre-loxP interaction in the major groove: Hint for structural distortion of mutant Cre and possible strategy for HIV-1 therapy

Kim

Lee

et al. 2000

J. Cell. Biochem.

Self Cite

View full text Add to dashboard Cite

Although the crystal structure of Cre recombinase complexed with DNA, named loxA, was elucidated a couple of years ago, it has not yet been determined which amino acids of the protein are involved in the specific Cre-loxP interaction. Arg259 and Gln90 interact with DNA substrate in the major groove from which the specificity of protein-DNA interaction comes. In this study, we substituted these residues for other amino acids. Also, two mutated DNA substrates were constructed. In each mutant, one of the bases that interact with Arg259 or Gln90 was changed into another base. In vitro binding assays and recombination assays of variant lox sites with wild-type and mutant-type Cre revealed that Arg259 plays a key role in Cre-loxP binding but Gln90 does not. However, the recombination activity still remained intact, although the binding between Cre and DNA substrate was not ensured.

show abstract

“…As proof of principle they used the Cre-lox system to reduce HIV replication in an in vitro model that introduced LoxP sites into the LTRs of the HIV (lox) laboratory strain of HIV. They found that Cre expression inhibited viral replication in HIV (lox)-infected cells and initiated genomic provirus excision, and this approach was subsequently extended by Lee et al (49, 50). This led to work from the groups of Buchholz and Hauber who were able to develop an evolved Cre recombinase that recognizes a loxP-like sequence found within the rare HIV isolate TZB0003.…”

Section: Introductionmentioning

confidence: 99%

Targeted gene disruption to cure HIV

Stone¹,

Kiem

Jerome

2013

Current Opinion in HIV and AIDS

View full text Add to dashboard Cite

Purpose of review Recent clinical research suggests that an HIV-infected patient with lymphoma who was transplanted with bone marrow homozygous for a disrupted mutant CCR5 allele has no remaining HIV replication and is effectively cured of HIV. Here we discuss approaches of disrupting host and viral genes involved in HIV replication and pathogenesis with the aim of curing patients with HIV. Recent findings Data from the ‘Berlin patient’ suggests that targeted gene disruption can lead to an HIV cure. This review discusses recent advances in the field of gene disruption towards the development of an anti-HIV therapy. We will introduce strategies to disrupt host and viral genes using precise disruptions, imprecise disruptions, or site-specific recombination. Furthermore, the production of engineered rare cutting endonucleases (zinc finger nucleases, TAL effector nucleases, and homing endonucleases) and recombinases that can recognize specific DNA target sequences and facilitate gene disruption will be discussed. Summary The discovery of a gene disruption approach that would cure or efficiently confine HIV infection could have broad implications for the treatment of millions of people infected with HIV. An efficient ‘one shot’ curative therapy would not only give infected patients hope of a drug- or treatment-free future, but could reduce the huge financial burden faced by many countries due to widespread administration of HAART.

show abstract

A Novel MutantloxP Containing Part of Long Terminal Repeat of HIV-1 in Spacer Region: Presentation of Possible Target Site for Antiviral Strategy Using Site-Specific Recombinase

Cited by 9 publications

References 19 publications

Chimeric recombinases with designed DNA sequence recognition

Chimeric recombinases with designed DNA sequence recognition

Characterization of Cre-loxP interaction in the major groove: Hint for structural distortion of mutant Cre and possible strategy for HIV-1 therapy

Targeted gene disruption to cure HIV

Contact Info

Product

Resources

About