HOME > product > Lifescience instrument > genome editing / Genome Editor Plus

Genome Editor Plus

Zygote Genome Editing, Tetraploid Formation, Oocyte Activation　

Genome Editor Plus ^NEW

This is ONE for all the early embryo experiment!

The function for oocyte activation and tetraploid formation is added to Genome Editor, which has been popular since it's released. This enables to do every experiments related to early embryos with only one unit of this Genome Editor plus.

Feature

The function to output AC voltage is added to Genome Editor, which enhanced zygote genome editing researches.

Two mode: AC + DC pulses for oocyte activation and tetraploid formation; and DC pulses for zygote genome editing.

Easy-to-use operation to program the parameter by touch panel

All the data of executed pulse output parameters can be exported through USB memory.

All the electrodes for CUY electroporator series and LF electrofusion series are connectable.

Application

High throughput and effcient CRISPR/Cas9-based mouse genome editing by RNA electroporation to zygote

Fgf10 homozygous mutant embryo have an easily detectable limbless phonotype. Electroporation of Cas9 mRNA (Cas9 Protein) and gRNA enables genome edit of Fgf10.

Electrode for zygote electroporation

LF501PT1-10 is applied for zygote genome editing, newly developed for zygoe electroporation.

Comparison between electroporation and microinjection

	Electroporation	Microinjection
Pre-operation	Not necessary	Preparation of injection and hold pippettes, etc.
Required time for 100 zygotes	~5 minutes	> 2 hours
Viability (after E15)	40-80%	10-50%
Efficiency	Same as microinjection	Depends on the sequence
Cost for devices	About $12,000	> $50,000
Required skills	Not necessary	Maniipularion of single zygotes
Required amount of Cas9 mRNA	500 - 2,000 ng	50 - 500 ng

More zygotes can be treated at one time by electroporation than microinjection.

Viability of treated zygotes are much higher in electroporation than in microinjection.

No special skills are required for electroporation compared to microinjection, which requires injection technique that is time-consuming to acquire.

Electroporation is , therefore, the ideal method for high throughput mouse zygote genome editing by CRISPR/Cas9 system.

References

1. Hashimoto M, Takemoto T. Electroporation enables the efficient mRNA delivery into the mouse zygotes and facilitates CRISPR/Cas9-based genome editing. Sci. Rep. 2015; 5: 11315.

2. Hashimoto M, Yamashita Y, Takemoto T. Electroporation of Cas9 protein/sgRNA into early pronuclear zygotes generates non-mosaic mutants in the mouse. Dev. Biol. 2016; 418(1); 1-9.

3. Tanihara F, Takemoto T, Kitagawa E, Rao S, Do LTK, Ohnishi A, et al. Somatic cell rprogramming-free generation of genetically modified pigs. Sci. Adv. 2016; 9(2); e1600803.

* The data above were obtained with CUY21EDIT II.

* The data is provided by Dr. Hashimoto, Osaka University, and Dr. Takemoto, Tokushima University.

Specification

GenomeEdit Mode

Voltage	1 - 200 V / 1 V resolution
Max current	1 A (1000 mA)
Pulse length (Pon)	0.10 - 1000 ms 0.10 - 9.99 ms (0.01 ms resolution) 10.0 - 99.9 ms (0.1 ms resolusion) 100 - 1000 ms (1 ms resolution)
Pulse interval	1 - 1000 ms 1.0 - 9.99 ms (0.01 ms resolution) 10.0 - 99.9 ms (0.1 ms resolution) 100 - 1000 ms (1 ms resolution)
Output pattern	Single polar pulse / Bi-polar pulse
Pulse number	1 - 1000 (for single polar pulse (Pd(+))） 1 - 500 (for Bi-polar pulse (Pd(+/-) or Pd(Alt))
Impedance measurement range	Max 4 kΩ

Fusion (Activation) Mode

AC Pulse

Voltage		1.0 - 20.0 V (0.1 V resolution)
Output time	0,1 - 99.9 sec (0.1 sec resolution）
Output time	Frequency	1 MHz

DC Pulse

Pulse Shape	Square pulse
Voltage	1 - 200 V (1 V resolusion)
Max current output	1 A (1,000 mA)
Pulse length	1 - 999 μ sec (1 μ sec resolution)
Pulse interval	0.01 - 9.99 sec (0.01 sec resolution)
Pulse number	1 - 1,000 pulses