Study of design parameters affecting the motion of DNA for nanoinjection

David, Regis A.; Jensen, Brian D.; Black, J.A.; Burnett, Sandra H.; Howell, Larry L.

doi:10.1088/0960-1317/22/5/055006

Cited by 4 publications

(2 citation statements)

References 33 publications

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“…A possible explanation for this behavior is that 4.5 mA protocols offers the best balance between effective electrical attraction/release of the DNA during the LAN process, while being a mild stressor in terms of cell viability. Even though the 1.5 mA protocol is milder in terms of cellular stress, a feature seen in electroporation studies to improve cell viability (Canatella et al 2001 ), perhaps the 4.5 mA protocol is better at balancing the cellular stress with effective attraction and release of the DNA, a parameter shown to increase DNA motion when done at higher magnitudes in processes like electrophoresis (David et al 2010 , 2011 , 2012 ).…”

Section: Discussionmentioning

confidence: 99%

CRISPR-Cas9 directed knock-out of a constitutively expressed gene using lance array nanoinjection

et al. 2016

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BackgroundCRISPR-Cas9 genome editing and labeling has emerged as an important tool in biologic research, particularly in regards to potential transgenic and gene therapy applications. Delivery of CRISPR-Cas9 plasmids to target cells is typically done by non-viral methods (chemical, physical, and/or electrical), which are limited by low transfection efficiencies or with viral vectors, which are limited by safety and restricted volume size. In this work, a non-viral transfection technology, named lance array nanoinjection (LAN), utilizes a microfabricated silicon chip to physically and electrically deliver genetic material to large numbers of target cells. To demonstrate its utility, we used the CRISPR-Cas9 system to edit the genome of isogenic cells. Two variables related to the LAN process were tested which include the magnitude of current used during plasmid attraction to the silicon lance array (1.5, 4.5, or 6.0 mA) and the number of times cells were injected (one or three times).ResultsResults indicate that most successful genome editing occurred after injecting three times at a current control setting of 4.5 mA, reaching a median level of 93.77 % modification. Furthermore, we found that genome editing using LAN follows a non-linear injection-dose response, meaning samples injected three times had modification rates as high as nearly 12 times analogously treated single injected samples.ConclusionsThese findings demonstrate the LAN’s ability to deliver genetic material to cells and indicate that successful alteration of the genome is influenced by a serial injection method as well as the electrical current settings.

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Section: Discussionmentioning

confidence: 99%

CRISPR-Cas9 directed knock-out of a constitutively expressed gene using lance array nanoinjection

et al. 2016

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“…Microinjection is also reported as being relatively effective and having low toxicity, but it is laborious and time intensive since cells are injected one at a time. Nanoinjection combines mechanical and electrical methods and has relatively high viability and efficiency [3][4][5][6][7][8]. However, the original nanoinjection system is also limited to single cell injections.…”

Section: Introductionmentioning

confidence: 99%

A microneedle array able to inject tens of thousands of cells simultaneously

Teichert¹,

Burnett²,

Jensen³

2013

J. Micromech. Microeng.

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This paper presents a biological microelectromechanical system for injecting foreign particles into thousands of cells simultaneously. The system inserts an array of microneedles into a monolayer of cells, and the foreign particles enter the cells by diffusion. The needle array is fabricated using a series of deep reactive ion etches and produces about 4 million needles that average 1 μm in diameter and 8 μm in length with 10 μm spacing. The insertion of the needles is controlled through a compliant suspension. The compliant suspension was designed to provide for needle motion into the cells while restraining rotations or transverse motions that could result in tearing of the cell membranes. Testing was performed using propidium iodide, a membrane impermeable dye, injected into HeLa cells. Average cell survivability was found to be 97.7%, and up to 97.9% of the surviving cells received the propidium iodide.

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Design and calibration of a six-axis MEMS sensor array for use in scoliosis correction surgery

Benfield

Yue

Lou

et al. 2014

J. Micromech. Microeng.

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A six-axis sensor array has been developed to quantify the 3D force and moment loads applied in scoliosis correction surgery. Initially this device was developed to be applied during scoliosis correction surgery and augmented onto existing surgical instrumentation, however, use as a general load sensor is also feasible. The development has included the design, microfabrication, deployment and calibration of a sensor array. The sensor array consists of four membrane devices, each containing piezoresistive sensing elements, generating a total of 16 differential voltage outputs. The calibration procedure has made use of a custom built load application frame, which allows quantified forces and moments to be applied and compared to the outputs from the sensor array. Linear or non-linear calibration equations are generated to convert the voltage outputs from the sensor array back into 3D force and moment information for display or analysis.

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Study of design parameters affecting the motion of DNA for nanoinjection

Cited by 4 publications

References 33 publications

CRISPR-Cas9 directed knock-out of a constitutively expressed gene using lance array nanoinjection

CRISPR-Cas9 directed knock-out of a constitutively expressed gene using lance array nanoinjection

A microneedle array able to inject tens of thousands of cells simultaneously

Design and calibration of a six-axis MEMS sensor array for use in scoliosis correction surgery

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