Lentiviral Manipulation of Gene Expression in Human Adult and Embryonic Stem Cells

Clements, Mark O.; Godfrey, A.; Crossley, Joanne; Wilson, Sam J.; Takeuchi, Asu; Boshoff, Chris

doi:10.1089/ten.2006.12.ft-95

Cited by 7 publications

(9 citation statements)

References 27 publications

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“…McMahon et al [78] have shown the superiority of lentiviral gene delivery to rat MSCs over other viral or nonviral gene delivery methods. Other studies reported the ability of lentiviral vectors to transduce human [79][80][81][82][83] or mouse [84] MSCs and sustain transgene expression longer than other viral and nonviral vectors. These studies have also indicated that the transduction procedure itself does not affect the growth, differentiation potential, and migration capacity of MSCs and does not induce excessive cell death.…”

Section: Genetic Manipulation Of Mscsmentioning

confidence: 99%

Introducing Transcription Factors to Multipotent Mesenchymal Stem Cells: Making Transdifferentiation Possible

2009

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Multipotent mesenchymal stem cells (MSCs) represent a promising autologous source for regenerative medicine. Because MSCs can be isolated from adult tissues, they represent an attractive cell source for autologous transplantation. A straightforward therapeutic strategy in the field of stem cell-based regenerative medicine is the transplantation of functional differentiated cells as cell replacement for the lost or defective cells affected by disease. However, this strategy requires the capacity to regulate stem cell differentiation toward the desired cell fate. This therapeutic approach assumes the capability to direct MSC differentiation toward diverse cell fates, including those outside the mesenchymal lineage, a process termed transdifferentiation. The capacity of MSCs to undergo functional transdifferentiation has been questioned over the years. Nonetheless, recent studies support that genetic manipulation can serve to promote transdifferentiation. Specifically, forced expression of certain transcription factors can lead to reprogramming and alter cell fate. Using such a method, fully differentiated lymphocytes have been reprogrammed to become macrophages and, remarkably, somatic cells have been reprogrammed to become embryonic stem-like cells. In this review, we discuss the past and current research aimed at transdifferentiating MSCs, a process with applications that could revolutionize regenerative medicine.

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Section: Genetic Manipulation Of Mscsmentioning

confidence: 99%

Introducing Transcription Factors to Multipotent Mesenchymal Stem Cells: Making Transdifferentiation Possible

2009

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“…The lentiviral vector G-P, which expresses short hairpin RNA (shRNA)-GFP, was described previously (9), and lentiviral vector X-P was generated using oligonucleotide 5Ј-CGA AAA AGA CTG CCA GAG ATC GAA AGA AGG CGA ACC TTC TCT CAA TCT CCG GCA GTC GGT GTT TCG TCC TTT CCA CAA GAT-3Ј and T7 oligonucleotide 5Ј-TAA TAC GAC TCA CTA TAG GG-3Ј to PCR amplify an shRNA expression cassette, using pGEM-U6L as a template (9). This amplicon was cloned using pGEM-T-Easy (Promega) and inserted into pCSPW as described previously (9).…”

Section: Rna Extraction and Reverse Transcriptase-pcr (Rt-pcr)mentioning

confidence: 99%

“…This amplicon was cloned using pGEM-T-Easy (Promega) and inserted into pCSPW as described previously (9). The shRNA sequences are therefore shRNA-GFP (5Ј-GUU CAU CUG CAC CAC CGG CAA GCU UCG GCU UGC CGG UGG UGC AGA UGA ACU U-3Ј) and shRNA-XBP)5Ј-GAC UGC CGG AGA UUG AGA GAA GGU UGC CCU UCU UUC GAU CUC UGG CAG UCU U-3Ј).…”

Section: Rna Extraction and Reverse Transcriptase-pcr (Rt-pcr)mentioning

confidence: 99%

X Box Binding Protein XBP-1s Transactivates the Kaposi's Sarcoma-Associated Herpesvirus (KSHV) ORF50 Promoter, Linking Plasma Cell Differentiation to KSHV Reactivation from Latency

Wilson¹,

Tsao²,

Webb³

et al. 2007

J Virol

Self Cite

101

130

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Reactivation of lytic replication from viral latency is a defining property of all herpesviruses. Despite this, the authentic physiological cues for the latent-lytic switch are unclear. Such cues should ensure that viral lytic replication occurs under physiological conditions, predominantly in sites which facilitate transmission to permissive uninfected cells and new susceptible hosts. Kaposi's sarcoma-associated herpesvirus (KSHV) is associated with the B-cell neoplasm primary effusion lymphoma (PEL), in which the virus remains latent. We have previously shown that PEL cells have the gene expression profile and immunophenotype of cycling preplasma cells (plasmablasts). Here, we show that the highly active spliced isoform of plasma cell transcription factor X box binding protein 1 (XBP-1s) is a lytic switch for KSHV. XBP-1s is normally absent in PEL, but the induction of endoplasmic reticulum stress leads to XBP-1s generation, plasma cell-like differentiation, and lytic reactivation of KSHV. XBP-1s binds to and activates the KSHV immediate-early gene ORF50 and synergizes with the ORF50 gene product RTA to induce a full lytic cycle. These data suggest that KSHV remains latent until B-cell terminal differentiation into plasma cells, the transcriptional environment of which provides the physiological "lytic switch" through XBP-1s. This links B-cell terminal differentiation to KSHV lytic reactivation.

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“…Generally, viral (e.g., adenoviruses, lentiviruses and retroviruses) and nonviral vectors (e.g., lipids and polymers) can be used for cellular transfection and/or nucleofection, thus offering durable gene expression within stem cells [93][94][95][96]. Nonviral carriers have a number of advantages over viral vectors, since they exhibit low-risk immunogenicity and insertional mutagenesis, controllable toxicity, and excellent gene-carrying capacity [95].…”

Section: A U T H O R P R O O Fmentioning

confidence: 99%

Nanodentistry: Combining Nanostructured Materials and Stem Cells for Dental Tissue Regeneration

2012

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Regenerative dentistry represents an attractive multidisciplinary therapeutic approach that complements traditional restorative/surgery techniques and benefits from recent advances in stem cell biology, molecular biology, genomics and proteomics. Materials science is important in such advances to move regenerative dentistry from the laboratory to the clinic. The design of novel nanostructured materials, such as biomimetic matrices and scaffolds for controlling cell fate and differentiation, and nanoparticles for diagnostics, imaging and targeted treatment, is needed. The combination of nanotechnology, which allows the creation of sophisticated materials with exquisite fine structural detail, and stem cell biology turns out to be increasingly useful in regenerative medicine. The administration to patients of dynamic biological agents comprising stem cells, bioactive scaffolds and/or nanoparticles will certainly increase the regenerative impact of dental pathological tissues. This overview briefly describes some of the actual benefits and future possibilities of nanomaterials in the emerging field of stem cell-based regenerative dentistry.

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Lentiviral Manipulation of Gene Expression in Human Adult and Embryonic Stem Cells

Cited by 7 publications

References 27 publications

Introducing Transcription Factors to Multipotent Mesenchymal Stem Cells: Making Transdifferentiation Possible

Introducing Transcription Factors to Multipotent Mesenchymal Stem Cells: Making Transdifferentiation Possible

X Box Binding Protein XBP-1s Transactivates the Kaposi's Sarcoma-Associated Herpesvirus (KSHV) ORF50 Promoter, Linking Plasma Cell Differentiation to KSHV Reactivation from Latency

Nanodentistry: Combining Nanostructured Materials and Stem Cells for Dental Tissue Regeneration

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