DIYGeneGun

SayedQuraishi

NilesNorthHighSchool
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TableofContents
Acknowledgements..3

Problem...4

ReviewofLiterature5

DesignPlan.....9

PerformanceCriteria..13

Data..14

DataAnalysis....15

Conclusion.17

References...18

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Acknowledgements
ThankyoutoMr.Eric,attheHomeDepotonHowardinEvanston,forgivingme

helpdevelopingmyprocedureforthegenegun.

ThankyoutoMs.Camelforsuggestinggenegunsasanareaofexplorationfor

myproject,aswellasbeingtheretosupportmyprojectthroughouttheyear.

Thankyoutomydadforlettingmeusehisofficetobuildthegenegun,and

makeitquiteabitmessy.

ThankyoutoDr.Bowman,fromtheUniversityofWyoming,forhelpingmeat

thebasisonhowtogothroughwithadesignprojectspecificallyinthemicrobiology

field.

ThankyoutoMr.Kyriazesforhelpingmecomeupwiththemateriallist,and

bringinguptheideatotheSIRSteachersatNilesNorth.

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Problem
Genegunsareanincrediblyscientificinstrument,anditsusesarereservedto

professorsatuniversitieswithlotsofscientificfunding.Theaveragegeneguncosts

around$20,000andthusitisquiteunrealisticforahighschoolorevensomesmall

collegestoobtainanduse.Genegunscouldbeusedtonotonlyeducate

microbiology,butalsodevelopnewcellstrainsforamultitudeofuses.Moreover,the

majorproblemisthatgenegunsarejusttooexpensivefortheaverageperson.The

purposeofthisdesignprojectwastoseeifitwaspossibletoproduceafunctioning

genegunatapricerangebelow$300.

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ReviewofLiterature
Agenegunmaysoundlikepartofasci-fimovieplot,butinrealityitisan

incrediblysoughtaftertoolinlaboratories.Agenegunisagunthatinessence

launchesDNAintoacellsnucleus.Genesareaunitofhereditaryinformationfrom

oneschromosomesandDNA.OncetheDNAenterthecellthroughbeingshotthrough

agenegun,itistransformedintoatransgene.Atransgeneisbasicallythemorphing

oftheDNAbeingshotandthecellbeingshotat(GeneGun,n.d).Ifthatisalittlehard

toimagine,thinkofbeingabletodesignacellthatcangoontoprocreate.The

transgenethencanrecreatethroughcellfunctionslikemitosiswhichprovidesa

uniquechancetomakeacombocell.

ThefirstgenegunwasproducedbyCornellProfessor,JohnSandfordandsome

graduatestudentsfrom1983-1986.Theyexperimentedwithonioncellsandtungsten

astheDNAbeingshotat.Theyusedaregular22calibernailgunandaimeditapetri

dishwithonioncells.Onioncellswerebeingusedbecauseoftheirlargesize,and

tungsten(chemicalelement)becauseofitsdensity.Afterafewtrialswerefinished

theyrealizedthatthegeneswereintertwiningandthatthissimpleideaactually

works(O'Brien,J.A.,&Lummis,S.C.Biolistictransfectionofneuronalculturesusing

ahand-heldgenegun2009).Thisthenbegancommercialgenegunproductionfor

useofitinlaboratories.Intodaysworldhowever,genegunsarenolongermadewith

riflesandinsteadmadewithextremelystronganddurablematerialtoensurethatthe
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genesremainpotentthroughoutthefiring,anditismucheasiertocheckand

examinepost-trial.

Genegunscanbeappliedinamultitudeofways.Genegunsmostpopularlyare

usedonplantcells.Theadvantagesofgenegunsbeingusedonplantcellsisthatit

canbeusedtomanipulatethebadplantcellsfromthegoodones(Bio-Rad

Laboratories,Inc.BiolisticParticleDeliverySystems.n.d).Forexample,ifaperson

hadanappleorchardandonlywantedbrightredappleswithnochanceofthem

becomingrotten,ageneguncouldpotentiallybemodifiedtoproduceaperfect

appleseedling.Genegunscanalsobeusedonhumansandanimals.Apopular

applicationofGenegunsforhumansareDNAvaccines.However,whilegengun

applicationsareveryuseful,genegunsareincrediblyexpensive.

Genegunsareveryexpensive.Theaveragecommercialgeneguncangofor

around$20,000.Foralotoflaboratories,thatisalotforascientifictooltheymay

onlyuseonceortwice.Duetothepricedilemmamanypeoplehavetriedtocomeup

withDIYguidestoproducetheirownhomemadegeneguns(HeliosGeneGun

System.n.d).Thehomemadegenegunsusuallylacktheprecisionoraccuracyofa

commercialgenegun,butcanhopefullycomeupwithasimilarrateofbeingableto

producetransgenes.Thecommercialgenegunwillfirewithanaverageof2/100cells

survivingandprocreating.Inotherwords,whilethemajorityofcellsdie,thelive

cellshavetheintendedimpactandwillbeevidentintheirabilitytobeusedinthe
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scientistsexperiment.Theyaresubstantiallycheaperwithitbeingonlyacouple

hundreddollarsincomparisontothethousandsofdollarsalegitimateonewould

cost(GlowingPlant.HowaGeneGunWorks.2013)..

Regardlessofhowitismade,commercialorhomemade,genegunshave

certainconstructsthatcannotbeignored.Theshootismadeupofusuallydense

elementssuchasgold,silver,andtungsten.Tungstenbecomesthemostcommon

elementbecauseofitscheapness.Tungstenhoweverdoesnotworkaswellasgoldor

silver.Whentungstenisshotatthecelltissue,itcanprovetobetoxictothecell

tissueandmaykillalotmorecellsthanprovideatransgene.Asfordiameterofthe

elementbeingshotitisusuallybetween1and4urn(ParticulateScienceand

Technology.2007).Thisisadjustablebasedonthesizeofthecellitisbeingshotat.

Thus,genegunshaveamoredistinctstrategicpointtheycanbeshotat.

Inconclusion,genegunsarematerialsthatcouldpotentiallybeincredibly

cheap.Genegunshaveproventobeasmartwaytotransformacell.Genegunscan

gothroughbiologicalprocessesandbeusedinamultitudeofpositiveways.Gene

gunscanbeusedtohelptheagriculturalissuesandmedicinalissuesfacingtheworld

today.Ifgenegunsaremoreaccessibletothepublicandnotrestrictedtoafew

scientistswhereitcannotbeseenbyneweyes,therecouldbealotmorewaystosee

itsuse.Hopefullythey'llbeavailabletothepublicatacheaperprice.Atsuchalow

priceitcouldbepotentiallybeusedinmiddleschools,highschools,andcollegeswith
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notaslargeofapublicendowment.Thus,thisiswhatIhopetodoinmydesign

project.

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DesignPlan
Belowisalogofchangesfromthebeginningofthedesignofthegenegun,uptillthe

endwhereIdidnotthinkIcouldmakeanybetterapplications.

ChangeinApplication Reason

3Pipe(width)basingwithan Thepipeatthatlengthissufficienttobe

attached4waycross thebaseofthemiddlesectionofthe

gun.Thecrossisattachedsotherestof

thegenegunspartscanbeadded.

Anotherpipe(width)wasattachedto Thiswastostemintothethinnerpipe

thecrossandthenanother3waycross thatwilllaterbeusedfortheprimary

wasadded genegunfunctions.3waycrosswasonly

neededbecauseitwouldntbeextended

anyfartherright

Avalvewasaddedthroughthecrossand Thepressuregaugeisnecessaryto

asmallerpipe administerthepressureofthegun,

specificallylaterwhentheCO2tire

inflatorisusedtomanagethelevels

beingbroughtintothecells.

Ontopofthevalvea1/12bushinwas Thisbushinwasaddedtomakeupfor
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addedtothepipeconnectingthevalve theCO2tireinflatorssmallandtight

openingwhichwouldntbeabletobe

screwedinwiththepipethere

AddtheCO2tireinflator TheCO2tireinflatoristhenaddedto

thebush,andcanbeusedtore

Addabushtothe3waycross Thisissotheotherpartsonthebottom

canbeadded

Fromthebushaddthepressuregauge Thepressuregaugeisaddedbecauseit

monitorsthepressurecominginfrom

theCO2inflator.Thiswilllaterbe

adjustedifnecessarytomakesurethe

gunisfiringcorrectly

Onthepipe,addacross Thiswillallowforfurtherattachments

necessarytofurtherthegoalsofthegun

Throughthecrossuseabushtothen Thiswilladdthesolenoidwhichcarries

insertthesolenoid theelectricalcurrentnecessaryforthe

guntofire

Thenaddthecontainertocrossatthe Thecontainerservesasthegenegun

bottomofthesolenoid. bulletglockwhichistheoverallpurpose
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ofthegun,andthisproject

Addawashertothebottomofthe Thisisforcontainmentpurposesaswell

container askeepingthebulletsinplace.The

bulletswilllaterbeputbyunscrewing

thewasherandinsertingthemin.

Screwinthesidewiththepipeintoa Thisisforthestabilityofthegunandso

squarefootofplywoodthatsathick thatitdoesntbreakorfallover

Use4screwsona4waybase

Useascrewgunotherwiseitwillbreak

Idealplacementofscrewingitinisdead theplywood

centeroftheboard

Atthispointscrewinthefiringswitchto Thefiringswitchatthecorneris

theleftcorneroftheboard. necessaryfortheelectricaloutlet

Addtherelaytothetopofthefiring Thisisforspaceconveniencesothatit

switch.Gluedtherelaytothetop hasaccesstothesameelectricityand

canbeaddedtothesolenoidlater

Connectthewiringofthesolenoidwith Thisissotheguncanbefired.

therelay.

Checkthefiring Makesureitfireswherethewasheris.It
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seemedtobeabitshaky

Adjustedthepressuretodifferentlevels Tomakesurethefiringswitchand

ofPSI everythingwasworkingasagunshould

GeneGunbulletsneedtobemade Genegunbulletsneededtobemadeata

cheapprice

Firstadministrationwasjusttotryit Theyweregrindeduptotrytomakea

out,andIusedtungstenshavingsand morehomogenousmixtureandsothat

onioncellsandgrindedthemup theywouldhaveahigherchanceofthem

interactingwhenthegunwasfired

AftergrindingthemupIusedwaxpaper Thisissoitwouldemulatebullets,and

towrapthemupandusedtapetoclose whatwasperceivedtokeepthebullets

offtheends alive

AfterthatIpluggedthegunintoa Thiswastoturnitonandsothegun

sourceintoa120Vplugoutlet couldfunction

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PerformanceCriteria
Thegunmustcostlessthan$300

Thefiringrangepressureneedstobealterable

Theelectricalaspectofthegunmustbeabletousedwithapowersourcethat

isworkablewith120Vconditions

ThegenesneedtointerchangetheDNA(betweenthetungstenandtheonion

cells)

Ifcellsdosurvivetheythenneedtobemonitoredtoseeiftheygothrough

biologicalfunctionslikemitosisandmeiosis.

Thegunitselfneedstobesafe,andbemonitoredsoitdoesntfireelsewhere

andhurtsomeone.

Totestifthegunisfunctionalisifitcanfire,buttoseeifthegenegunworks

thecombinedcellsneedtogrowandsurvive.

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Data

LIveTungstenCells LiveOnionCells LiveCells

Trial1 0 0 0

Trial2 0 0 0

Trial3 0 0 0

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DataAnalysis
ThedatashowsthatthisDIYgenegundidnotfunctionashowitwasintended

too.Allthecellsdiedthroughoutthe3trials.Aftertryingtofixtherelayandthe

pressureitwasatmultipletimes(redesign&retreat),evenatthebareminimum

neededforittofire,theshotendedupkillingallthecellsinthebullet.Thequestion

thoughishowisitsurethatthecellsdiediftheyaresosmall.Theanswerwas

obviousbecausethetungstenwasfromgrayscrewshavings,andwellonionsareatan

color,andeverytrialresultedinablackalmostashysubstance,butseparatedfrom

deadonioncells(whichwerealittlelighter)thanthedeadtungstencells.
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Thedatamostlikelyshowsthattherewasanerrorinthegenegunbulletsor

thegenegunbulletcompartment.Thusshouldtherehavebeentimetogobackand

researchhowtomakeastrongerbulletcompartmentandbulletsthatfunctionwith

thecellstohopefullymakeaconjoinedcell.Also,maybetherewasanissuewiththe

powerofthegun,andinordertofixthattherelaywouldneedtobeadjusted.

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Conclusion

Inconclusion,whilethegenegundidnotworkasintended,anddidnotfollow

alloftheperformancecriteria,thisprojectwasntnecessarilyafailure.Thegene

gunsmajorerrorwasthatthefiringresultedindeadcells.Withthatinformationit

suggeststhatthegunpartdidworkandtherewassomethingwrongwithhowthe

bulletsweremade,and/orthegunwastoostrong.Theperformancecriteriaofit

beingrelativelyinexpensiveatapricebelow$300wasfollowed,andprettymuch

everythingwasfollowedbutthegenesbecomingconjoinedandworking.

Moreover,theimpactofthisprojectshowsthatthisissomethingtotallydoable

withmoretime.Overallthoughtheimpactofthisprojectisimmense.Shouldthe

bulletsbemadeproperlyandthecompartmentisadjustedandadaptedtothe

situation,thesecheapgunscouldhaveahugeeffectoneducationandscientific

discoveries.AsmentionedpriorlyaDIYgeneguncouldbemadeandusedinmiddle

schools,highschools,andevencollegestonotonlyteachmicrobiology,andbiology,

butalsousedforexperiments.Itcouldbeusedinscientificexperimentsofall

scientistsandeveninpoorerpartsofthecountryandtheworld.

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References
WorksCited:

Bio-RadLaboratories,Inc.BiolisticParticleDeliverySystems.Bulletin

5443.http://www.bio-rad.com/webroot/web/pdf/lsr/literature/Bulletin_5443.p

df.

GeneGun.(n.d.).Retrievedfrom

http://passel.unl.edu/pages/informationmodule.php?idinformationmodule=95807

7244&topicorder=3&maxto=7

GlowingPlant.HowaGeneGunWorks.YouTube,publishedSeptember

27,2013.https://www.youtube.com/watch?v=VqklR_8YRfA.

HeliosGeneGunSystem.(n.d.).RetrievedNovember07,2016,from

http://www.bio-rad.com/en-cn/product/helios-gene-gun-system

O'Brien,J.A.,&Lummis,S.C.(2009,March02).Biolistictransfectionofneuronalcultures

usingahand-heldgenegun.RetrievedNovember07,2016,from

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2649370/

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ParticulateScienceandTechnology.(2007,April25).RetrievedNovember07,2016,from

http://www.tandfonline.com/doi/abs/10.1080/02726358708904533?src=recsys