Optical delivery of multiple opsin-encoding genes leads to targeted expression and white-light activation

Kamal Dhakal; Subrata Batabyal; Weldon Wright; Young-tae Kim; Samarendra Mohanty

doi:10.1038/lsa.2015.125

Go to Nature Website

Your Location：

Home >

Browse articles >

Optical delivery of multiple opsin-encoding genes leads to targeted expression and white-light activation

Original Articles

- Optical delivery of multiple opsin-encoding genes leads to targeted expression and white-light activation
- Light: Science & Applications Vol. 4, Issue 4, (2015)
- Affiliations：
  
  1.Biophysics and Physiology Lab, The University of Texas at Arlington, Arlington, TX, USA
  2.NanoScope Technologies, LLC, Arlington, TX, USA
  3.Department of Bioengineering, The University of Texas at Arlington, Arlington, TX, USA
- Author bio：
  
  S Mohanty, Email: smohanty@uta.edu
- Funds：
- DOI：10.1038/lsa.2015.125
  CLC：
- Published：2015-12，
  
  Published Online：07 July 2015，
  
  Received：23 December 2014，
  
  Revised：30 June 2015，
  
  Accepted：04 July 2015
- Accepted：
Scan QR Code
Dhakal, K. et al. Optical delivery of multiple opsin-encoding genes leads to targeted expression and white-light activation. Light: Science & Applications, 4, e352 (2015).
DOI：

Dhakal, K. et al. Optical delivery of multiple opsin-encoding genes leads to targeted expression and white-light activation. Light: Science & Applications, 4, e352 (2015). DOI： 10.1038/lsa.2015.125.

摘要

Abstract

In photodegenerative diseases such as retinitis pigmentosa (RP) and age-related macular degeneration (AMD)

progressive loss of vision occurs as a result of degeneration of the periphery of the retina and the macula

respectively. Current optogenetic stimulation-based approaches to vision restoration offer the advantages of cellular specificity

high resolution

and minimal invasiveness over electrode arrays; however

the clinical translation of optogenetic activation suffers from the lack of a method for the delivery of opsins into spatially targeted regions of a retina that has degenerated. Non-targeted opsin delivery through viral or non-viral methods to non-photodegenerated retinal areas will perturb these already functioning retinal regions. Furthermore

viral methods are subject to limitations on the delivery of large plasmids

such as fusion constructs of multiple spectrally separated opsins (e.g.

channelrhodopsin-2 (ChR2)

chimeric opsin variants (C1V1)

ReaChR)

which can provide higher photo-excitability than can a single narrow-band opsin under ambient light conditions. Here

we report the ultrafast near-infrared laser-based spatially targeted transfection of single and multiple opsins and present a comparison with the opsin expression distribution achieved using another non-viral

but non-targeted

transfection method

lipofection. Functional evaluation of cells transfected with multiple opsins using the laser method revealed a significantly higher white-light-induced photocurrent than in cells expressing a single opsin (ChR2). The laser-assisted targeted delivery of multiple opsin-encoding genes to the peripheral retina/macula is ideal for sensitizing retinal areas that have degenerated

thus paving the way toward the restoration of lost vision in RP/AMD patients.

关键词

Keywords

gene deliveryoptogeneticsoptoporationvision restoration

references

Bi AD, Cui JJ, Ma YP, Olshevskaya E, Pu MLet al. Ectopic expression of a microbial-type rhodopsin restores visual responses in mice with photoreceptor degeneration.Neuron2006;50: 23–33..

Thyagarajan S, van Wyk M, Lehmann K, Löwel S, Feng GPet al. Visual function in mice with photoreceptor degeneration and transgenic expression of channelrhodopsin 2 in ganglion cells.J Neurosci2010;30: 8745–8758..

Zhang Y, Ivanova E, Bi AD, Pan ZH. Ectopic expression of multiple microbial rhodopsins restores ON and OFF light responses in retinas with photoreceptor degeneration.J Neurosci2009;29: 9186–9196..

Tomita H, Sugano E, Isago H, Hiroi T, Wang Zet al.Channelrhodopsin-2gene transduced into retinal ganglion cells restores functional vision in genetically blind rats.Exp Eye Res2010;90: 429–436..

Lagali PS, Balya D, Awatramani GB, Münch TA, Kim DSet al. Light-activated channels targeted to ON bipolar cells restore visual function in retinal degeneration.Nat Neurosci2008;11: 667–675..

Busskamp V, Duebel J, Balya D, Fradot M, Viney TJet al. Genetic reactivation of cone photoreceptors restores visual responses in retinitis pigmentosa.Science2010;329: 413–417..

Boyden ES, Zhang F, Bamberg E, Nagel G, Deisseroth K. Millisecond-timescale, genetically targeted optical control of neural activity.Nat Neurosci2005;8: 1263–1268..

Nagel G, Szellas T, Huhn W, Kateriya S, Adeishvili Net al. Channelrhodopsin-2, a directly light-gated cation-selective membrane channel.Proc Natl Acad Sci USA2003;100: 13940–13945..

Cao H, Gu L, Mohanty SK, Chiao JJ. An Integrated μLED Optrode for Optogenetic Stimulation and Electrical Recording.IEEE Trans Biomed Eng2013;60: 225–229..

Dhakal KR, Gu L, Shivalingaiah S, Dennis TS, Morris-Bobzean SAet al. Non-scanning fiber-optic near-infraredbeam led to two-photon optogenetic stimulation in-vivo.PLoS ONE2014;9: e111488..

Zhang F, Aravanis AM, Adamantidis A, de Lecea L, Deisseroth K. Circuit-breakers: optical technologies for probing neural signals and systems.Nat Rev Neurosci2007;8: 577–581..

Mohanty SK, Reinscheid RK, Liu X, Okamura N, Krasieva TBet al. In-depth activation of Channelrhodopsin 2-sensitized excitable cells with high spatial resolution using two-photon excitation with a near-infrared laser microbeam.Biophys J2008;95: 3916–3926..

Satpathy S, Batabyal S, Dhakal KR, Lin J, Kim Yet al. Broad spectral excitation of opsin for enhanced stimulation of cells.Opt Lett2015;40: 2465–2468..

Mohanty SK, Lakshminarayananan V. Optical techniques in optogenetics.J Modern Optics2015;62: 949–970..

Tirlapur UK, König K. Targeted transfection by femtosecond laser.Nature2002;418: 290–291..

Soman P, Zhang WD, Umeda A, Zhang ZJ, Chen SC. Femtosecond laser-assisted optoporation for drug and gene delivery into single mammalian cells.J Biomed Nanotechnol2011;7: 334–341..

Stracke F, Rieman I, König K. Optical nanoinjection of macromolecules into vital cells.J Photoch Photobio B2005;81: 136–142..

Barrett LE, Sul JY, Takano H, van Bockstaele EJ, Haydon PGet al. Region-directed phototransfection reveals the functional significance of a dendritically synthesized transcription factor.Nat Methods2006;3: 455–460..

Peng C, Palazzo RE, Wilke I. Laser intensity dependence of femtosecond near-infrared optoinjection.Phys Rev E2007;75: 041903..

Kohli V, Elezzabi AY. Laser surgery of zebrafish (Danio rerio) embryos using femtosecond laser pulses: optimal parameters for exogenous material delivery, and the laser's effect on short- and long-term development.BMC Biotechnol2008;8: 7..

Lei M, Xu HP, Yang H, Yao BL. Femtosecond laser-assisted microinjection into living neurons.J Neurosci Meth2008;174: 215–218..

Uchugonova A, König K, Bueckle R, Isemann A, Tempea G. Targeted transfection of stem cells with sub-20 femtosecond laser pulses.Opt Express2008;16: 9357–9364..

Dhakal K, Black B, Mohanty S. Introduction of impermeable actin-staining molecules to mammalian cells by optoporation.Sci Rep2014;4: 6553..

Zeira E, Manevitch A, Khatchatouriants A, Pappo O, Hyam Eet al. Femtosecond infrared laser – an efficient and safein vivogene delivery system for prolonged expression.Mol Ther2003;8: 342–350..

Gu L, Mohanty SK. Targeted microinjection into cells and retina using optoporation.J Biomed Opt2011;16: 128003..

Buj R, Iglesias N, Planas AM, Santalucía T. A plasmid toolkit for cloning chimeric cDNAs encoding customized fusion proteins into any Gateway destination expression vector.BMC Mol Biol2013;14: 18..

Kong X, Mohanty SK, Stephens J, Heale JT, Gomez-Godinez Vet al. Comparative analysis of different laser systems to study cellular responses to DNA damage in mammalian cells.Nucleic Acids Res2009;37: e68..

Chakravarty P, Qian W, El-Sayed MA, Prausnitz MR. Delivery of molecules into cells using carbon nanoparticles activated by femtosecond laser pulses.Nat Nanotechnol2010;5: 607–611..

Gu L, Koymen AR, Mohanty SK. Crystalline magnetic carbon nanoparticle assisted photothermal delivery into cells using CW near-infrared laser beam.Sci Rep2014;4: 5106..

Xiong RH, Raemdonck K, Peynshaert K, Lentacker I, de Cock Iet al. Comparison of gold nanoparticle mediated photoporation: vapor nanobubbles outperform direct heating for delivering macromolecules in live cells.Acs Nano2014;8: 6288–6296..

Lukianova-Hleb EY, Mutonga MBG, Lapotko DO. Cell-specific multifunctional processing of heterogeneous cell systems in a single laser pulse treatment.ACS Nano2012;6: 10973–10981..

Views

Downloads

CSCD

Alert me when the article has been cited

Submit

Tools

Publicity Resources

Wide-field three-photon excitation in biological samples

Correction-free remotely scanned two-photon in vivo mouse retinal imaging

Related Author

Christopher J Rowlands

Demian Park

Oliver T Bruns

Kiryl D Piatkevich

Dai Fukumura

Rakesh K Jain

Moungi G Bawendi

Edward S Boyden

Related Institution

Department of Biological Engineering, Massachusetts Institute of Technology

Media Lab, Massachusetts Institute of Technology

Department of Chemistry, Massachusetts Institute of Technology

Edwin L. Steele Laboratories, Department of Radiation Oncology, Massachusetts General Hospital and Harvard Medical School

Department of Brain and Cognitive Sciences, McGovern Institute and MIT Center for Neurobiological Engineering, Massachusetts Institute of Technology

⁰