**5. References**

Abrahams, P. J. & Van Der Eb, A. J. (1975). In vitro transformation of rat and mouse cells by DNA from simian virus 40. *J Virol*, 16, 1, pp. 206-209

Al-Dosari, M. S. & Gao, X. (2009). Nonviral Gene Delivery: Principle, Limitations, and Recent Progress. *Aaps Journal*, 11, 4, pp. 671-681

**Sample composition Zeta potential (mV)** 

Table 6. Surface charge of Sr and Mg substituted calcium phosphate nanoparticles

Nano-particulate calcium phosphate has shown several interesting advantages in biomedical applications because of its biocompatibility and easy preparation process. The DNA condensation characteristic of nano-particulate calcium phosphate makes it a potential choice for gene therapy system applications. Nano-particulate calcium phosphates are able to condense DNA strands, carry them in the blood, deliver the genetic material to target

Therefore, there is a common agreement among most of the works regarding gene delivery application on utilizing the calcium phosphate to deliver the gene into the nucleus; the final target of gene therapy methods. Because of the advantages of the DNA/calcium phosphate complex, it is one of the highly appealing systems currently studied, although it has been used in in-vitro gene delivery for many years already. The translation of its application into

Researchers need to solve the instability of calcium phosphate in physiological conditions. If calcium phosphate/DNA complexes degrade in the blood circuit, it cannot be used in most of the clinical gene delivery applications. The other problem is the low transfection efficiency, which currently limits the application of the system. There are controversial reports about the transfection efficiency of calcium phosphate/DNA system, mostly because of instability and the complicated nature of calcium phosphate in solution. Once these problems are overcome by adequate novel technologies, the excellent biocompatibility and

Abrahams, P. J. & Van Der Eb, A. J. (1975). In vitro transformation of rat and mouse cells by

Al-Dosari, M. S. & Gao, X. (2009). Nonviral Gene Delivery: Principle, Limitations, and Recent

0.0Sr-CaP 4.5±0.1 0.5Sr-CaP 5.0±0.2 1.0Sr-CaP 6.1±0.1 5.0Sr-CaP 7.3±0.3 10.0Sr-CaP 7.8±0.2

0.0Mg-CaP 3.2±0.5 0.25Mg-CaP 6.7±0.4 0.50Mg-CaP 7.5±1 1.0Mg-CaP 8±0.8

(Reproduce from (Hanifi et al., 2010a; Hanifi et al., 2010b)).

cells, and move them into cells resulting in reasonable transcription.

biodegradability of calcium phosphate remains as a major advantage.

DNA from simian virus 40. *J Virol*, 16, 1, pp. 206-209

Progress. *Aaps Journal*, 11, 4, pp. 671-681

**Sr-Substituted CaP** 

**Mg-Substituted CaP** 

clinical therapy methods requires more work.

**4. Conclusion** 

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**25** 

Hugo Peluffo

*Uruguay* 

**Modular Multifunctional Protein** 

*Department of Histology and Embryology, Faculty of Medicine, University of the Republic* 

The introduction of genes into the organism or the regulation of the expression of endogenous genes has emerged in the last decade as a very potent strategy for correcting monogenic inherited diseases, treating acute disorders, and slowing down the progression of diseases without known cure. In addition it constitutes an important tool for research, which has been widely used and has contributed to show the mechanisms behind several

Adequate carriers able to transfer DNA or RNA into target cells have been largely explored. However, this is an area under continuous expansion as there is no ideal vector suitable for all applications. In fact, no individual vector will meet all the characteristics for a perfect or ideal vector, as many of the needs are different and even contradictory. For example, immunogenicity is in most cases an undesirable side effect, while it is a valuable property when treating tumours as it contributes to their clearance. Another example of contradictory needs of one single vector would be the capacity of a vector to determine the overexpression of the transgenic protein for life. This would be an essential property for the treatment of inherited diseases produced by the lack of a particular protein, however for the treatment of acute injuries the lifelong expression of a therapeutic protein will probably be deleterious. Moreover, some vectors do not transduce post-mitotic cells like neurons or muscle fibres, which is a drawback for targeting these cell types but may be an advantage for the targeting of cancer cells. Thus, there is a need for diverse type of vectors for diverse therapeutic or experimental paradigms, and in particular versatile tuneable vectors would be very interesting. Moreover, several basic problems with the known vectors persist, like toxicity, oncogenicity, immunogenicity, low transfection efficiency, or poor bioavailability, which

Due to their natural efficiency, viruses have been modified to act as vectors, and they have shown a good degree of success. Non-viral vectors have also been developed by combining several properties necessary for transfection: nucleic acid attachment and condensation, cell attachment, cell entry, endosomal escape, intracellular trafficking, nuclear entry, and nucleic acid release. Some of these vectors are quite simple, as the ones formed by the combination of nucleic acids and lipid components or other carriers like polyethylene glycol (PEG). Others include the previous components but have in addition attached targeting molecules like antibodies, enabling these vectors to preferentially transfect a given tissue. In fact even

**1. Introduction** 

physiological processes and pathologies.

need further consideration and efforts.

*(UDELAR) and Neurodegeneration Laboratory, Institut Pasteur de Montevideo* 

**Vectors for Gene Therapy** 

