Electroporation is a highly efficient delivery method for transformation in various cell types; however, in microalgae there is lack of optimized electroporation parameters due to cell wall, protoplast preparation and viability. Therefore, we optimized electroporation conditions for transforming microalgae using Chlamydomonas reinhardtii strains of wild type and mutant (cell wall deficient). To investigate the effects of molecule size, calcein (623 Da) and fluorescein isothiocyanate-dextran (FITC-dextran, 40 kDa) were used and various electroporation parameters were applied such as different voltage and pulse length and molecule uptake pattern and cell viability were observed. Cell wall is insignificant in case of small sized molecule uptake as noticed by 1.25 kV/cm and 30 ms for both strains, whereas for larger molecules by 1.5 and 2 kV/cm and 30 ms for mutant and wild type, respectively. In terms of viability, there was no significant difference in both the strains on applied electroporation parameters. The controlled parameters corresponding to 1.5 to 2.0 kV/cm and 20 to 30 ms could be used to deliver macromolecules (DNA, proteins) into cells effectively.

Electroporation Microalgae Chlamydomonas reinhardtii Molecules Delivery Viability

Inderwildi OR, King DA, Energy Environ. Sci., 2, 343 (2009)
Lu J, Sheahan C, Fu P, Energy Environ. Sci., 4, 2451 (2011)
Mata TM, Martins AA, Caetano NS, Renewable and Sustainable Energy Reviews., 14, 217 (2010)
Chisti Y, Biotechnol. Adv., 25, 294 (2007)
Beer LL, Boyd ES, Peters JW, Posewitz MC, Curr. Opin.Biotechnol., 20, 264 (2009)
Davis SWDSC, Boundy RG, Ed., Transportation energy data book, the Center for Transportation Analysis of the Oak Ridge National Laboratory (2012)
Demirbas A, Energy Policy, 35(9), 4661 (2007)
Knothe G, Sharp CA, Ryan TW, Energy Fuels, 20(1), 403 (2006)
Radakovits R, Jinkerson RE, Darzins A, Posewitz MC, Eukaryot Cell., 9, 486 (2010)
Qin S, Lin H, Jiang P, Biotechnol. Adv., 30, 1602 (2012)
Harris EH, Annual Review of Plant Physiology and Plant Molecular Biology., 52, 363 (2001)
Hoober JK, Science., 246, 1503 (1989)
Shrager J, Hauser C, Chang CW, Harris EH, Davies J, McDermott J, Tamse R, Zhang Z, Grossman AR, Plant Physiol., 131, 401 (2003)
Grossman AR, Croft M, Gladyshev VN, Merchant SS, Posewitz MC, Prochnik S, Spalding MH, Curr. Opin. Plant Biol., 10, 190 (2007)
Rochaix JD, Vandillewijn J, Nature., 296, 70 (1982)
Neumann E, Schaeferridder M, Wang Y, Hofschneider PH, Embo. J., 1, 841 (1982)
Zimmermann U, Pilwat G, Riemann F, Biochimica Et Biophysica Acta., 375, 209 (1975)
Gehl J, Acta Physiologica Scandinavica., 177, 437 (2003)
Brown LE, Sprecher SL, Keller LR, Molecular and Cellular Biology., 11, 2328 (1991)
Shimogawara K, Fujiwara S, Grossman A, Usuda U, Genetics., 148, 1821 (1998)
Canatella PJ, Karr JF, Petros JA, Prausnitz MR, Biophysical J., 80, 755 (2001)
Azencott HR, Peter GF, Prausnitz MR, Ultrasound in Medicine and Biology., 33, 1805 (2007)
Yang C, Owen HA, Yang P, J. Cell. Biol., 180, 403 (2008)
Lelong A, Hegaret H, Soudant P, Research in Microbiology., 162, 969 (2011)
Kolber Z, Falkowski PG, Limnology and Oceanography., 38, 1646 (1993)
Richard JP, Melikov K, Vives E, Ramos C, Verbeure B, Gait MJ, Chernomordik LV, Lebleu B, J. Biological Chem., 278, 585 (2003)
Sun Y, Yang ZY, Gao XS, Li QY, Zhang QQ, Xu ZK, Molecular Biotechnology., 30, 185 (2005)
Kilian O, Benemann CSE, Niyogi KK, Vick B, Proceedings of the National Academy of Sciences of the United States of America., 108, 21265 (2011)
Weaver JC, Chizmadzhev YA, Bioelectrochemistry and Bioenergetics., 41, 135 (1996)
Monk BC, Adair WS, Cohen RA, Goodenough UW, Planta., 158, 517 (1983)