Baculoviral transduction facilitates TALEN-mediated targeted transgene integration and Cre/LoxP cassette exchange in human-induced pluripotent stem cells

Zhu, Hongxia; Lau, Cia-Hin; Goh, Sal-Lee; Liang, Qingle; Chen, Can; Du, Shouhui; Phang, Rui-Zhe; Tay, Felix Chang; Tan, Wee-Kiat; Li, Zhendong; Tay, Johan Chin-Kang; Fan, Weimin; Wang, Shu

doi:10.1093/nar/gkt721

Cited by 50 publications

(38 citation statements)

References 39 publications

(47 reference statements)

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“…Since LV predominantly integrates into transcriptional active regions in the host genome [4,33,34] BVs belong to the family of Baculoviridae and exhibit a doublestranded circular DNA genome in a filamentous shape. They primarily infect insects but BV derived vectors are also capable of infecting various mammalian cell types including also human embryonic stem cells [38] and pluripotent stem cells [39,40]. BV vectors are derived from the well characterized Autographa californica multiple nucleopolyhedrovirus (AcMNPV), which contains a genome of 134 kb with 155 genes.…”

Section: Viral Vectors For Talen Deliverymentioning

confidence: 99%

“…To date there are two studies demonstrating that viral vectors can be explored for delivery of TALEN expression cassettes into target cells [40,52]. In the first study lentiviral and adenoviral vectors and their ability to result in sufficient expression of functional TALENs in the desired target cells was investigated.…”

Section: Molecular Design and Functionality Of Talen Delivering Viralmentioning

confidence: 99%

“…However, there is evidence that reverse transcriptase (RT) of simple and complex retroviruses possesses low affinity to their RNA template and therefore switching events within the template during the reverse transcription reaction from RNA to DNA may occur [54][55][56]. Due to the fact that the presence of direct repeats increases these switching events resulting in deletion of one or more repeats [54][55][56][57], it seems that the usage of commonly used versions of LV vector systems for sufficient delivery and expression of TALENs in mammalian cells is not feasible.In the second study a BV vector system was used to deliver TALENs into U87 glioblastoma-derived cells and human-induced pluripotent stem cells (h-iPSCs) [40]. Unlike the study by Holkers and colleagues [58], this study was based on a single vector containing both TALEN expression cassettes in one virus genome (BV-TALEN) (Figure 2).…”

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confidence: 99%

“…It could also be demonstrated that transduction of BV-TALEN at moderate MOI resulted in no genomic instability or increased cell death in glioma cells. After co-transduction of U87 cells with BV-TALEN and a BV vector containing an neomycin-eGFP-expression cassette (Neo-eGFP) flanked by homologous sequences to the AAVS1 locus, Zhu et al [40] could show stable integration of the Neo-eGFP cassette into the AAVS1 locus. Efficiency was determined by measuringe GFP expression levels, a PCR approach detecting the modified AAVS1 locus, and Southern blot analysis, reaching a value of up to 95% eGFP positive cells.…”

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confidence: 99%

“…Furthermore, activity of the TALEN pair at the targeted chromosomal location in combination with unspecific binding and activity of the same TALEN pair at off-target sites may also lead to chromosomal translocations by linking the two translocation breakpoints. Although there is no direct evidence that chromosomal translocations can be induced after introducing a single TALEN pair into mammalian cells, there is evidence that translocations can be specifically induced in the presence of two TALEN pairs targeting different loci within the host genome [59,60].To circumvent these problems it could be beneficial to design viral vectors that contain both expression cassettes of a TALEN pair in one construct as conducted in the BV-based study [40]. This approach may decrease toxic side effects induced by the viral vector itself, because a decreased viral dose may be required for sufficient TALEN delivery.…”

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Progress and Problems with Viral Vectors for Delivery of Talens

Bergmann¹

2014

J Mol Genet Med

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Transcription Activator-Like Effector Nucleases (TALENS) and Their Mode of ActionFor sequence-specific genome engineering and its biotechnological and medical applications various systems were tested. Most recent tools include designer nucleases and the bacteria derived clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 system using short RNA to induce precise cleavage at endogenous genomic loci [1,2]. Designer nucleases are mainly represented by zinc finger nucleases (ZFNs) and transcription activator-like effector nucleases (TALENs) [3][4][5][6][7], combining features of a customized DNA binding motif for sequence-specific DNA binding domain and a nuclease for introduction of doubled-strand DNA (dsDNA) breaks at the target site.TALENs are a promising new class of designer nucleases that can be specifically designed to bind DNA sequences of interest and to introduce dsDNA breaks. They are chimeric proteins consisting of a N-terminal nuclear localization signal, a central DNA binding domain and a C-terminal FokI nuclease domain. The DNA binding domain originates from transcription activator like effectors (TALEs) of the bacterial plant pathogen Xanthomonas, used by this bacterium to alter the expression of several host genes [8]. Its main characteristic is a central repeat region consisting of a variable number of incomplete tandem repeats that are usually comprised of 33-35 amino acids (aa) that are identical except for the hypervariable repeat-variable diresidue (RVD) at a positions 12 and 13 [6]. With some degree of degeneracy, the RVD of each repeat is specific for binding a corresponding nucleotide in their contiguous target DNA sequence [6,8]. The C-terminal FokI domain provides non-specific endonuclease activity and similar to ZFNs, binding of the TALE domains upstream and downstream of the respective DNA target and subsequent dimerization at the FokI nuclease domain lead to dsDNA breaks. It is of note that the spacer between the TALEN DNA binding sites needs to be considered when designing novel TALENs [6,7]. After binding, dsDNA breaks are introduced which can than activate cellular pathways either leading to homologous recombination in the presence of the respective homologous donor AbstractIt has long been envisaged that gene disruption or gene correction in affected target cells can be efficiently conducted in vitro and in vivo and over the recent years several tools for achieving this goal were developed. Designer nucleases such as zinc finger nucleases (ZFNs) were extensively explored and more recently transcription activator-like effector nucleases (TALENs) were introduced for sequence-specific genome engineering in the mammalian genome. ZFNs and TALENs are fusion proteins containing a customized DNA-binding motif for sequence-specific DNA binding linked to a nuclease for introduction of double-stranded DNA breaks. Both systems were explored in mammalian cells using non-viral and viral delivery methods. Herein, we will provide a state-ofthe-art overview of available virus-base...

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Section: Viral Vectors For Talen Deliverymentioning

confidence: 99%

Section: Molecular Design and Functionality Of Talen Delivering Viralmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Progress and Problems with Viral Vectors for Delivery of Talens

Bergmann¹

2014

J Mol Genet Med

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Cartilage Tissue Engineering: Recent Advances and Perspectives from Gene Regulation/Therapy

2015

Adv Healthcare Materials

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Diseases in articular cartilages affect millions of people. Despite the relatively simple biochemical and cellular composition of articular cartilages, the self-repair ability of cartilage is limited. Successful cartilage tissue engineering requires intricately coordinated interactions between matrerials, cells, biological factors, and phycial/mechanical factors, and still faces a multitude of challenges. This article presents an overview of the cartilage biology, current treatments, recent advances in the materials, biological factors, and cells used in cartilage tissue engineering/regeneration, with strong emphasis on the perspectives of gene regulation (e.g., microRNA) and gene therapy.

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