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Review
. 2020 Dec 11;12(12):1427.
doi: 10.3390/v12121427.

Toward Tightly Tuned Gene Expression Following Lentiviral Vector Transduction

Audrey Page  1 , Floriane Fusil  1 , François-Loïc Cosset  1
Affiliations
Review

Toward Tightly Tuned Gene Expression Following Lentiviral Vector Transduction

Audrey Page et al. Viruses. .

Abstract

Lentiviral vectors are versatile tools for gene delivery purposes. While in the earlier versions of retroviral vectors, transgene expression was controlled by the long terminal repeats (LTRs), the latter generations of vectors, including those derived from lentiviruses, incorporate internal constitutive or regulated promoters in order to regulate transgene expression. This allows to temporally and/or quantitatively control transgene expression, which is required for many applications such as for clinical applications, when transgene expression is required in specific tissues and at a specific timing. Here we review the main systems that have been developed for transgene regulated expression following lentiviral gene transfer. First, the induction of gene expression can be triggered either by external or by internal cues. Indeed, these regulated vector systems may harbor promoters inducible by exogenous stimuli, such as small molecules (e.g., antibiotics) or temperature variations, offering the possibility to tune rapidly transgene expression in case of adverse events. Second, expression can be indirectly adjusted by playing on inserted sequence copies, for instance by gene excision. Finally, synthetic networks can be developed to sense specific endogenous signals and trigger defined responses after information processing. Regulatable lentiviral vectors (LV)-mediated transgene expression systems have been widely used in basic research to uncover gene functions or to temporally reprogram cells. Clinical applications are also under development to induce therapeutic molecule secretion or to implement safety switches. Such regulatable approaches are currently focusing much attention and will benefit from the development of other technologies in order to launch autonomously controlled systems.

Keywords: induction; integration; lentiviral vectors; promoter; sensor; signal; synthetic biology; transgene.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Externally inducible promoter principle. (a) The TET-system. For the tet-ON version, upon addition of doxycycline, the tetracycline controlled trans-activator (tTA) can no longer bind Tet response element (TRE), hence leading to interruption of gene expression, whereas for the tet-OFF version, reverse tetracycline controlled trans-activator (rtTA) is not bound to TRE in absence of doxycycline, which does not allow gene transcription until addition of doxycycline. Several strategies have been tested for implantation of the Tet-system in lentiviral vectors (LVs). Initially, two vectors, one encoding the target gene under the control of a TRE and another encoding the trans-activator or the trans-repressor were used. These two parts were then combined into a single vector. Moving to a step further to reduce background level, the trans-activator/trans-repressor were also placed under the regulation of TRE. (b) The LoxP/Cre system. For this system, two versions have also been tested in LVs. The gene of interest can be placed between two LoxP sites. Consequently, upon Cre exposure, it will be excised and hence stop transgene expression. Alternatively, a gene silencing sequence (stop codon, poly(A) signal) is inserted with LoxP sites around, just before the target gene. Consequently, the addition of Cre recombinase will lead to the removal of the stop codon and lead to transgene expression.
Figure 2
Figure 2
Synthetic biology approaches. Lentiviral vectors are used to transduce cells. These vectors encode the main parts of the synthetic network: receptors, transducers, and effectors. Inputs signals that are markers of a pathological condition will be sensed by specific membranous or intracellular receptors, inducing a transducing cascade. This signal integration will lead to the expression of effector molecules such as shRNA, cytokines, or pro-apoptotic cues.
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References

    1. Frecha C., Costa C., Lévy C., Nègre D., Russell S.J., Maisner A., Salles G., Peng K.-W., Cosset F.-L., Verhoeyen E. Efficient and Stable Transduction of Resting B Lymphocytes and Primary Chronic Lymphocyte Leukemia Cells Using Measles Virus gp Displaying Lentiviral Vectors. Blood. 2009;114:3173–3180. doi: 10.1182/blood-2009-05-220798. - DOI - PubMed
    1. Girard-Gagnepain A., Amirache F., Costa C., Lévy C., Frecha C., Fusil F., Nègre D., Lavillette D., Cosset F.-L., Verhoeyen E. Baboon Envelope Pseudotyped LVs Outperform VSV-G-LVs for Gene Transfer into Early-Cytokine-Stimulated and Resting HSCs. Blood. 2014;124:1221–1231. doi: 10.1182/blood-2014-02-558163. - DOI - PubMed
    1. Sandrin V., Boson B., Salmon P., Gay W., Nègre D., Le Grand R., Trono D., Cosset F.-L. Lentiviral Vectors Pseudotyped with a Modified RD114 Envelope Glycoprotein Show Increased Stability in Sera and Augmented Transduction of Primary Lymphocytes and CD34+ Cells Derived from Human and Nonhuman Primates. Blood. 2002;100:823–832. doi: 10.1182/blood-2001-11-0042. - DOI - PubMed
    1. Duvergé A., Negroni M. Pseudotyping Lentiviral Vectors: When the Clothes Make the Virus. Viruses. 2020;12:1311. doi: 10.3390/v12111311. - DOI - PMC - PubMed
    1. Frank A.M., Braun A.H., Scheib L., Agarwal S., Schneider I.C., Fusil F., Perian S., Sahin U., Thalheimer F.B., Verhoeyen E., et al. Combining T-cell-specific Activation and In Vivo Gene Delivery Through CD3-Targeted Lentiviral Vectors. Blood Adv. 2020;4:5702–5715. doi: 10.1182/bloodadvances.2020002229. - DOI - PMC - PubMed

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