Electroporation Protocols Microorganism, Mammalian System, and Nanodevice

This third edition provides in-depth knowledge on the delivery of naked DNA and small-interfering RNA (siRNA) to the targeted microorganism, mammalian single cells, tissues, and animals for prevention and treatment of disease. It builds on the success of the first edition and on the progress made in siRNA delivery and DNA vaccines for large animals as well as discovery of electroporation applications for the fragile tissues and for internal organs. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

 Authoritative and easily accessible, Electroporation Protocols: Microorganism, Mammalian System, and Nanodevice, Third Edition aims to be an invaluable resource for investigators both in and outside of this field. 

Part I:  Design and Fabrication of Micro-/Nano-Electroporation

1. Cell Size-specific Transfection by Micropillar Array Electroporation

Xuan Liu, Yingbo Zu, and Shengnian Wang

2. Soft Electroporation through 3D Hollow Nanoelectrodes

Jun Yin and Yang Li

Lei Li

Lingqian Chang, Stephen Black, Chandani Chitrakar, and Mehdi Nouri

5. Low-Voltage Flow-Through Electroporation Membrane and Method

Juliette Experton, Aaron G. Wilson, and Charles R. Martin

Section II: Micro-/Nano-electroporation for in vitro Delivery

6. Nanofountain Probe Electroporation for Monoclonal Cell Line Generation

Horacio D. Espinosa, Prithvijit Mukherjee, and Cesar Patino

7. 3D Nanochannel Electroporation for Macromolecular Nucleotides Delivery

Lingqian Chang, Chandani Chitrakar, and Mehdi Nouri

8. Nanoelectroporation and Collection of Genetically Modified Exosomes in Primary Cultures of Dendritic Cells

Silvia Duarte-Sanmiguel, Natalia Higuita-Castro, and Daniel Gallego-Perez

9. Electroporation of CRISPR-Cas9 into Malignant B Cells for Loss-of-function Studies of Target Gene via Knockout

Jian Yu

10. Microfluidic Device for Localized Electroporation

Part III: Advanced in vivo Electroporation

11. Transdermal Delivery of Nucleic Acid Mediated by Punching and Electroporation

Dong Huang, Yuanyu Huang, and Zhihong Li

12. Targeted in vivo Electroporation Using Nanoengineered Microelectrodes

Daniel Schwarz and Andreas T. Schaefer

13. Zygote Electroporation for CRISPR/Cas9 Delivery to Generate Genetically Modified Mice

Tatsuya Takemoto

14. Genome Editing in Staphylococcus aureus by Conditional Recombineering and CRISPR/Cas9-mediated Counterselection

Kelsi Penewit and Stephen J. Salipante

15. Tissue Nano-Transfection of Surgically Exposed Stroma

Section IV: Microorganism Electroporation

16. Transformation of a Model Microalga, Chlamydomonas reinhardtii without Cell-wall Removal

Takashi Yamano and Hideya Fukuzawa

17. Electroporation Transformation Protocol for Phaeodactylum tricornutum

Hanhua Hu and Yufang Pan

18. Efficient Transformation of the Diatoms Phaeodactylum tricornutum by Multi-pulse Electroporation

Kentaro Ifuku and Dongyi Yan

19. Rapid and Effective Electroporation Protocol for Nannochloropsis oceanica

20. Electroporation Procedures for Genetic Modification of Green Algae Chlorella spp.

21. Electrotransformation of Saccharomyces cerevisiae

Meilhoc E and Teissié J