Novel Cell and Gene Therapies for HIV

Hoxie, James A.; June, Carl H.

doi:10.1101/cshperspect.a007179

Cited by 56 publications

(63 citation statements)

References 153 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Linkage of artificial and natural gene control systems will be useful in the construction of effective and safe systematic gene therapy protocols and transgenic organisms (43). For example, many genes or RNAs have been tested for gene therapy for HIV (46,47). The gene transfer and expression systems are ready in CD4 ϩ T cells (48,49).…”

Section: Discussionmentioning

confidence: 99%

Inducible Suppression of Global Translation by Overuse of Rare Codons

Kobayashi

2015

Appl Environ Microbiol

View full text Add to dashboard Cite

(7). In many cases, artificial gene networks have focused on switching gene expression for specific purposes by using the fluorescent protein gene as an output gene (1, 2). The control of cell behavior with artificial gene networks requires an output gene whose biochemical function is clearly defined and a deficient mutant form thereof (4). To extend the application of artificial gene networks, we need a more convenient gene as the output gene in order to control biological activity in various organisms.The present study focused on codon usage bias, and an artificial gene-silencing system was constructed that is different from the repressor protein or interfering RNA found in a natural biological system. Low-usage codons have a genome-wide distribution at a very low frequency, and their tRNAs occur in the cell at lower concentrations than those for normal codons (8-10). The expression of foreign genes is suppressed in Escherichia coli because of differences in codon usage (11,12). Overexpression of the integrase mRNA containing 20 rare arginine codons with a strong ribosome binding site reduces the growth rate of the host E. coli (11). Therefore, the pathway of a rare codon in the genome and its corresponding tRNA is one "vulnerability" or "security hole" of biological systems, much like one in a computer system or network (13,14). In the present study, I constructed an artificial gene containing many low-usage codons to exploit this security hole by monopolizing multiple minor codon tRNAs through its expression, resulting in the suspension of almost all translation in the cell (Fig. 1). This scheme was modeled on a type of computer system or network attack known as a distributed denial-of-service attack. I arranged this artificial gene downstream of the lac promoter of E. coli, the galactose-inducible promoter of yeast, and the doxycycline (Dox)-inducible promoter of mammalian cells. I found repression of the growth of E. coli, yeast, and mammalian cells, as well as downregulation of gene expression with the induction of my artificial gene. The application of this artificial gene provides a novel nonspecific virus defense system in E. coli and human cells. This artificial gfp gene would work as a system device with which to control cell behavior with an artificial gene network with applications in biotechnology. MATERIALS AND METHODS Plasmid construction, genes, cells, phages, and chemicals. (i) E. coli andSaccharomyces cerevisiae. The artificial green fluorescent protein (GFP) genes lgfp and hgfp were designed on the basis of the codon usage database (15) and synthesized by the GenScript Corporation (Piscataway, NJ). The C-terminal deletion mutant genes lgfp⌬1, lgfp⌬2, and lgfp⌬3, lacking 25, 50, and 75% of the length of the C terminus of lgfp, respectively, were obtained by PCR from the lgfp gene (Table 1). The codon adaptation indices (CAIs) of artificial genes were calculated with the formula (16), f(i) and f(j) are frequencies of synonymous codons for amino acids, and L is the number of codons.All plasmid...

show abstract

Section: Discussionmentioning

confidence: 99%

Inducible Suppression of Global Translation by Overuse of Rare Codons

Kobayashi

2015

Appl Environ Microbiol

View full text Add to dashboard Cite

show abstract

“…An ideal therapy for HIV or other chronic viral infections that course with latent reservoirs is believed to involve the generation of a source of long-lived, self-renewing, and multilineage hematopoi- etic stem cells that repopulate the host with genetically modified cells that are refractory to infection (45,46). Since the unique and exceptional case of an HIV-1 sterilizing cure of a patient due to bone marrow transplantation with a matched donor homozygote for the CCR5⌬32 mutation (47,48), alternative strategies have aimed to reproduce the CCR5⌬32 phenotype using genome-editing tools.…”

Section: Discussionmentioning

confidence: 99%

Zinc Finger Endonuclease Targeting PSIP1 Inhibits HIV-1 Integration

Badía

Pauls

Riveira-Muñoz

et al. 2014

Antimicrob Agents Chemother

View full text Add to dashboard Cite

Genome editing using zinc finger nucleases (ZFNs) has been successfully applied to disrupt CCR5 or CXCR4 host factors and inhibit viral entry and infection. Gene therapy using ZFNs to modify the PSIP1 gene, which encodes the lens epithelium-derived growth factor (LEDGF) protein, might restrain an early step of the viral replication cycle at the integration level. ZFNs targeting the PSIP1 gene (ZFN LEDGF ) were designed to specifically recognize the sequence after the integrase binding domain (IBD) of the LEDGF/p75 protein. ZFN LEDGF successfully recognized the target region of the PSIP1 gene in TZM-bl cells by heteroduplex formation and DNA sequence analysis. Gene editing induced a frameshift of the coding region and resulted in the abolishment of LEDGF expression at the mRNA and protein levels. Functional assays revealed that infection with the HIV-1 R5 BaL or X4 NL4-3 viral strains was impaired in LEDGF/p75 knockout cells regardless of entry tropism due to a blockade in HIV-1 proviral integration into the host genome. However, residual infection was detected in the LEDGF knockout cells. Indeed, LEDGF knockout restriction was overcome at a high multiplicity of infection, suggesting alternative mechanisms for HIV-1 genome integration rather than through LEDGF/p75. However, the observed residual integration was sensitive to the integrase inhibitor raltegravir. These results demonstrate that the described ZFN LEDGF effectively targets the PSIP1 gene, which is involved in the early steps of the viral replication cycle; thus, ZFN LEDGF may become a potential antiviral agent for restricting HIV-1 integration. Moreover, LEDGF knockout cells represent a potent tool for elucidating the role of HIV integration cofactors in virus replication.

show abstract

“…As the cells made resilient by CCR5 alteration alone remain vulnerable to CXCR4 tropic viruses and the reverse also holds good, C46 can be effectively used to inhibit infection with both of the viral strains [152]. Extraneously administered genetic material coding for dominant negative inhibitory proteins of HIV replication such as the M-10 and those coding for intrabodies and intrakines can also cause favourable intracellular modifications in the Tcells making them resistant to HIV infection [148].…”

Section: Dna Manipulationmentioning

confidence: 99%

“…Modification of mature CD4 T-cells confers protection only during their life span and warrant the need for repetitive transfusions over time. On the other hand, genetically modified stem cells such as the CD34+ haematopoietic stem cells can be effective with a single transplantation as exemplified with replacement therapy [148].…”

Section: Dna Manipulationmentioning

confidence: 99%