Nucleofection

<h2 id="applications">Applications</h2>
<p>Nucleofection is a method to transfer substrates into <a href="/facts/Mammal/phSwTzBo">mammalian cells</a> so far considered difficult or even impossible to transfect. Examples for such substrates are nucleic acids, like the DNA of an isolated <a href="/facts/Gene/e1swwPY6">gene</a> <a href="/facts/Molecular_cloning/Te56373e">cloned</a> into a <a href="/facts/Plasmid/yf59JVlA">plasmid</a>, or <a href="/facts/Small_interfering_RNA/891ytkD9">small interfering RNA</a> (siRNA) for knocking down expression of a specific <a href="/facts/Endogenous/DFkSLkRI">endogenous</a> gene.
</p><p>Primary cells, for example <a href="/facts/Stem_cell/qgB9dFrI">stem cells</a>, especially fall into this category, although many other <a href="/facts/Cell_culture/7Qhu13D9">cell lines</a> are also difficult to transfect. Primary cells are freshly isolated from <a href="/facts/Tissue_(biology)/H7CpML7A">body tissue</a> and thus cells are unchanged, closely resembling the <a href="/facts/In-vivo/kWcWyl6f">in-vivo</a> situation, and are therefore of particular relevance for <a href="/facts/Biomedical_research/lSVBFxDJ">medical research</a> purposes. In contrast, cell lines have often been cultured for decades and may significantly differ from their origin.
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<h2 id="mechanism">Mechanism</h2>
<p>Based on the physical method of <a href="/facts/Electroporation/Dx7tt65o">electroporation</a>, nucleofection uses a combination of electrical parameters, generated by a device called Nucleofector, with cell-type specific reagents. The substrate is transferred directly into the <a href="/facts/Cell_nucleus/D7sFRa3c">cell nucleus</a> and the <a href="/facts/Cytoplasm/Wa0sE6xf">cytoplasm</a>. In contrast, other commonly used non-viral transfection methods rely on <a href="/facts/Cell_division/n5MrunWj">cell division</a> for the transfer of DNA into the nucleus. Thus, nucleofection provides the ability to transfect even non-dividing cells, such as <a href="/facts/Neurons/G7CyTVdH">neuron</a> and resting <a href="/facts/Blood_cells/lf9GA0Dj">blood cells</a>. Before the introduction of the Nucleofector Technology, efficient <a href="/facts/Horizontal_gene_transfer/FPyUIsf2">gene transfer</a> into primary cells had been restricted to the use of <a href="/facts/Viral_vector/sqkIdcqo">viral vectors</a>, which typically involve disadvantages such as safety risks, lack of reliability, and high cost. The non-viral gene transfer methods available were not suitable for the efficient transfection of primary cells. Non-viral delivery methods may require cell division for completion of transfection, since the DNA enters the nucleus during breakdown of the <a href="/facts/Nuclear_envelope/IwRQ5BVL">nuclear envelope</a> upon <a href="/facts/Cell_division/n5MrunWj">cell division</a> or by a specific localization sequence.
Optimal nucleofection conditions depend upon the individual <a href="/facts/List_of_distinct_cell_types_in_the_adult_human_body/IjWXx31l">cell type</a>, not on the substrate being transfected. This means that identical conditions are used for the nucleofection of DNA, RNA, siRNAs, <a href="/facts/ShRNA/RAqm3gpI">shRNAs</a>, <a href="/facts/MRNA/oUL6qxQn">mRNAs</a> and <a href="/facts/Precursor_mRNA/4UXtKPHw">pre-mRNAs</a>, <a href="/facts/Bacterial_artificial_chromosome/1MzCeAoe">BACs</a>, <a href="/facts/Peptide/C4ltc7UG">peptides</a>, <a href="/facts/Morpholino/EsU9Heor">morpholinos</a>, <a href="/facts/Peptide_nucleic_acid/9MkVaqXK">PNA</a>, or other biologically active molecules.
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<h2 id="see-also">See also</h2>
<ul><li><a href="/facts/Electroporation/Dx7tt65o">Electroporation</a></li></ul>

<ul><li>Kerima Maasho; Alina Marusina; Nicole M. Reynolds; John E. Coligan; Francisco Borrego (January 2004). "Efficient gene transfer into the human natural killer cell line, NKL, using the amaxa nucleofection system". <i>Journal of Immunological Methods</i>. 284 (1–2): 133–140. <a href="/facts/Doi_(identifier)/muM9Etpq">doi</a>:<a href="https://doi.org/10.1016%2Fj.jim.2003.10.010">10.1016/j.jim.2003.10.010</a>. <a href="/facts/PMID_(identifier)/JlHAvMHt">PMID</a> <a href="https://pubmed.ncbi.nlm.nih.gov/14736423">14736423</a>.</li>
<li>Pascal G. Leclere; Aliza Panjwani; Reginald Docherty; Martin Berry; John Pizzey; David A. Tonge (15 Mar 2005). "Effective gene delivery to adult neurons by a modified form of electroporation". <i>Journal of Neuroscience Methods</i>. 142 (1): 137–143. <a href="/facts/Doi_(identifier)/muM9Etpq">doi</a>:<a href="https://doi.org/10.1016%2Fj.jneumeth.2004.08.012">10.1016/j.jneumeth.2004.08.012</a>. <a href="/facts/PMID_(identifier)/JlHAvMHt">PMID</a> <a href="https://pubmed.ncbi.nlm.nih.gov/15652627">15652627</a>. <a href="/facts/S2CID_(identifier)/ldJsHa2Y">S2CID</a> <a href="https://api.semanticscholar.org/CorpusID:38463232">38463232</a>.</li>
<li>Michela Aluigi; Miriam Fogli; Antonio Curti; Alessandro Isidori; Elisa Gruppioni; Claudia Chiodoni; Mario P. Colombo; Piera Versura; Antonia D'Errico-Grigioni; Elisa Ferri; Michele Baccarani; Roberto M. Lemoli (February 2006). <a href="https://doi.org/10.1634%2Fstemcells.2005-0198">"Nucleofection is an efficient non-viral transfection technique for human bone marrow-derived mesenchymal stem cells"</a>. <i>Stem Cells</i>. 24 (2): 454–461. <a href="/facts/Doi_(identifier)/muM9Etpq">doi</a>:<a href="https://doi.org/10.1634%2Fstemcells.2005-0198">10.1634/stemcells.2005-0198</a>. <a href="/facts/PMID_(identifier)/JlHAvMHt">PMID</a> <a href="https://pubmed.ncbi.nlm.nih.gov/16099993">16099993</a>.</li></ul>

Nucleofection open-in-new

Nucleofection