384:
multiple templates of the DNA construct is a distinct possibility, resulting in potential variable expression levels and copy numbers of the inserted gene. This is due to the ability of the constructs to give and take genetic material from other constructs, causing some to carry no transgene and others to carry multiple copies; the number of copies inserted depends on both how many copies of the transgene an inserted construct has, and how many were inserted. Also, because eukaryotic constructs rely on
79:
20:
269:
If the delivered DNA construct contains a selectable marker, then stably transformed cells can be selected and cultured using tissue culture methods. For example, if the delivered DNA construct contains a gene that confers resistance to an antibiotic or herbicide, then stably transformed cells may be
173:
Biolistic transformation involves the integration of a functional fragment of DNA—known as a DNA construct—into target cells. A gene construct is a DNA cassette containing all required regulatory elements for proper expression within the target organism. While gene constructs may vary in their design
149:
below. The bullet welded to the disk below the Petri plate, and the genetic material blasted into the sample with a doughnut effect involving devastation in the middle of the sample with a ring of good transformation around the periphery. The gun was connected to a vacuum pump and was placed under a
383:
Biolistics introduces DNA randomly into the target cells. Thus the DNA may be transformed into whatever genomes are present in the cell, be they nuclear, mitochondrial, plasmid or any others, in any combination, though proper construct design may mitigate this. The delivery and integration of
217:
A gene encoding a selectable marker is a common element within DNA constructs and is used to select for properly transformed cells. The selectable marker chosen will depend on the species being transformed, but it will typically be a gene granting cells a detoxification capacity for certain
193:
Promoters control the location and magnitude of gene expression and function as “the steering wheel and gas pedal” of a gene. Promoters precede the gene of interest in the DNA construct and can be changed through laboratory design to fine-tune transgene expression. The 35S promoter from
261:
of undifferentiated plant cells or a group of immature embryos growing on gel medium in a Petri dish. After the DNA-coated gold particles have been delivered to the cells, the DNA is used as a template for transcription (transient expression) and sometimes it integrates into a plant
59:
with a gene of interest and firing these micro-projectiles into cells using mechanical force, an integration of desired genetic information can be introduced into desired cells. The technique involved with such micro-projectile delivery of DNA is often referred to as
322:
The gene gun has become a common tool for labeling subsets of cells in cultured tissue. In addition to being able to transfect cells with DNA plasmids coding for fluorescent proteins, the gene gun can be adapted to deliver a wide variety of vital dyes to cells.
366:
a transgene inserted with a gene gun. Additionally, with only one firing of a gene gun, a skilled technician can generate two transformed organisms in certain species. This technology has even allowed for modification of specific tissues
204:
Terminator sequences are required for proper gene expression and are placed after the coding region of the gene of interest within the DNA construct. A common terminator for biolistic transformation is the NOS terminator derived from
240:
sequences that allow for controlled removal of the construct from the target genome. Such elements are chosen by the construct developer to perform specialized functions alongside the main gene of interest.
314:
The delivery of plasmids into rat neurons through the use of a gene gun, specifically DRG neurons, is also used as a pharmacological precursor in studying the effects of neurodegenerative diseases such as
211:. Due to the high frequency of use of this terminator in genetically engineered plants, strategies have been developed to detect its presence within the food supply to monitor for unauthorized GE crops.
92:
The pressure eventually reaches the point where the rupture disk breaks, and the resulting burst of helium propels the DNA/gold-coated macrocarrier ('Plastic Disk') into the stopping screen.
840:"Glyphosate Resistance as a Novel Select-Agent-Compliant, Non-Antibiotic-Selectable Marker in Chromosomal Mutagenesis of the Essential Genes asd and dapB of Burkholderia pseudomallei"
134:
which would relay a signal if proper insertion of the DNA transcript occurred. Genetic transformation was demonstrated upon observed expression of the marker gene within onion cells.
82:
A gene gun is used for delivery of exogenous DNA to cells. This method is known as 'biolistics'. Gene guns can be used effectively on most cells but are mainly used on plant cells.
285:. The new plant that originated from a successfully transformed cell may have new traits that are heritable. The use of the gene gun may be contrasted with the use of
960:
Carrer, Helaine; Hockenberry, Tish Noel; Svab, Zora; Maliga, Pal (October 1993). "Kanamycin resistance as a selectable marker for plastid transformation in tobacco".
362:
mediated transformation, which have difficulty targeting the vector to and stably expressing in the chloroplast. In addition, there are no reports of a chloroplast
145:
cylinder (bullet) down a 22 caliber
Douglas barrel. A droplet of the tungsten powder coated with genetic material was placed onto the bullet and shot down into a
632:
150:
vacuum while firing. The early design was put into limited production by a Rumsey-Loomis (a local machine shop then at
Mecklenburg Road in Ithaca, NY, USA).
67:
This device is able to transform almost any type of cell and is not limited to the transformation of the nucleus; it can also transform organelles, including
358:
is a product of biolistics. Plastid transformation has also seen great success with particle bombardment when compared to other current techniques, such as
281:, and each may divide and differentiate into the organized, specialized, tissue cells of an entire plant. This capability of total re-generation is called
347:
Biolistics has proven to be a versatile method of genetic modification and it is generally preferred to engineer transformation-resistant crops, such as
130:
between 1983 and 1986. The original target was onions (chosen for their large cell size), and the device was used to deliver particles coated with a
1389:
561:
Sanford, J.C.; Klein, T.M.; Wolf, E.D.; Allen, N. (1987). "Delivery of substances into cells and tissues using a particle bombardment process".
525:
1327:
157:
as a non-explosive propellant and a multi-disk collision delivery mechanism to minimize damage to sample tissues. Other heavy metals such as
588:
Klein, T.M.; Wolf, E.D.; Wu, R.; Sanford, J.C. (May 1987). "High-velocity microprojectiles for delivering nucleic acids into living cells".
755:
Benfey, P. N.; Chua, N.-H. (1990-11-16). "The
Cauliflower Mosaic Virus 35S Promoter: Combinatorial Regulation of Transcription in Plants".
165:
are also used to deliver genetic material with gold being favored due to lower cytotoxicity in comparison to tungsten projectile carriers.
1962:
95:
When the macrocarrier hits the stopping screen, the DNA-coated gold particles are propelled through the screen and into the target cells.
2045:
415:"Nano-biolistics: A method of biolistic transfection of cells and tissues using a gene gun with novel nanometer-sized projectiles"
352:
1835:
1174:
Kikkert, Julie; Vidal, Jose; Reisch, Bruce (2005). "Stable
Transformation of Plant Cells by Particle Bombardment/Biolistics".
153:
Biolistics, Inc sold Dupont the rights to manufacture and distribute an updated device with improvements including the use of
1191:
1120:
739:
473:
2051:
2041:
1254:"Modifications to the hand-held Gene Gun: improvements for in vitro Biolistic transfection of organotypic neuronal tissue"
1992:
249:
Gene guns are mostly used with plant cells. However, there is much potential use in humans and other animals as well.
2002:
1320:
1232:
1674:
174:
depending on the desired outcome of the transformation procedure, all constructs typically contain a combination a
905:"Hygromycin B phosphotransferase as a selectable marker for DNA transfer experiments with higher eucaryotic cells"
2010:
1553:
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392:, they cannot be targeted to specific locations within the genome, unless the transgene is co-delivered with
198:
is an example of a commonly used promoter that results in robust constitutive gene expression within plants.
1528:
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1313:
388:—a process by which the transgene is integrated into the genome without similar genetic sequences—and not
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1007:
Gilbertson, Larry (December 2003). "Cre–lox recombination: Cre-ative tools for plant biotechnology".
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536:
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195:
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331:
316:
1103:
Praitis, Vida (2006). "Creation of
Transgenic Lines Using Microparticle Bombardment Methods".
1443:
258:
237:
179:
1982:
1757:
1523:
851:
764:
597:
137:
The earliest custom manufactured gene guns (fabricated by Nelson Allen) used a 22 caliber
8:
1972:
1904:
1842:
1706:
1336:
1056:"Multicolor "DiOlistic" Labeling of the Nervous System Using Lipophilic Dye Combinations"
660:"Physical Trauma and Tungsten Toxicity Reduce the Efficiency of Biolistic Transformation"
175:
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613:
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464:
Carter, Matt; Shieh, Jennifer (6 March 2015). "Chapter 11 - Gene
Delivery Strategies".
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selected by including that antibiotic or herbicide in the tissue culture media.
1935:
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1987:
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635:. Cornell University College of Agriculture and Life Sciences. Archived from
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Norris, M. H.; Kang, Y.; Lu, D.; Wilcox, B. A.; Hoang, T. T. (2009-07-31).
792:
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56:
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Transformed cells can be treated with a series of plant hormones, such as
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Helium fills the chamber and pressure builds against the rupture disk.
2115:
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1957:
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1543:
1502:
1484:
609:
219:
44:
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O'Brien, J; Holt, M; Whiteside, G; Lummis, SC; Hastings, MH (2001).
2105:
2072:
1875:
1870:
138:
115:
68:
1147:
Sanford, John (April 28, 2006). "Biolistic plant transformation".
734:(2 ed.). Oxford, New York, USA: Oxford University Press Inc.
2100:
2077:
1657:
1407:
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1374:
658:
Russell, Julie A.; Roy, Mihir K.; Sanford, John C. (1992-03-01).
348:
111:
108:
52:
1107:. Methods in Molecular Biology. Vol. 351. pp. 93–108.
236:
Optional components of a DNA construct include elements such as
1178:. Methods in Molecular Biology. Vol. 286. pp. 61–78.
1142:
1140:
371:, although this is likely to damage large numbers of cells and
274:
162:
154:
127:
1251:
299:
for different methods of transformation in different species.
1167:
1137:
959:
489:
Sanford, John C. (1990). "Biolistic plant transformation".
158:
78:
19:
48:
40:
1222:
732:
1227:. Springer Science & Business Media. p. 131.
1049:
902:
815:"nopaline synthase terminator: Topics by Science.gov"
560:
1390:
List of varieties of genetically modified maize/corn
1223:
Hayward, M.D.; Bosemark, N.O.; Romagosa, T. (2012).
1050:
Gan, Wen-Biao; Grutzendler, Jaime; Wong, Wai Thong;
730:Slater, Adrian; Scott, Nigel; Fowler, Mark (2008).
1173:
837:
587:
526:"Biologist invent gun for shooting cells with DNA"
729:
657:
412:
2128:
903:Blochlinger, K; Diggelmann, H (December 1984).
725:
723:
721:
719:
717:
715:
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523:
517:
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1963:Detection of genetically modified organisms
630:
463:
413:O'Brien, John A.; Lummis, Sarah CR (2011).
326:Gene gun bombardment has also been used to
302:
1328:
1314:
1006:
754:
624:
554:
1071:
936:
879:
691:
440:
430:
307:Gene guns have also been used to deliver
1300:John O'Brien presents...Gene Gun Barrels
1225:Plant Breeding: Principles and Prospects
581:
375:, rather than all, cells of the tissue.
86:The gene gun apparatus is ready to fire.
77:
18:
1335:
1146:
1102:
488:
406:
2129:
844:Applied and Environmental Microbiology
182:sequence, the gene of interest, and a
39:is a device used to deliver exogenous
1309:
1302:for more information about biolistics
64:, short for "biological ballistics".
962:MGG Molecular & General Genetics
295:to insert DNA into plant cells. See
257:The target of a gene gun is often a
55:to cells. By coating particles of a
23:PDS-1000/He Particle Delivery System
13:
1993:Genetic use restriction technology
1245:
1161:10.1111/j.1399-3054.1990.tb05888.x
563:Particulate Science and Technology
503:10.1111/j.1399-3054.1990.tb05888.x
218:herbicides or antibiotics such as
102:
37:biolistic particle delivery system
16:Device used in genetic engineering
14:
2173:
1293:
2011:Cartagena Protocol on Biosafety
1258:Journal of Neuroscience Methods
1216:
1096:
1043:
1000:
953:
896:
831:
807:
748:
524:Segelken, Roger (14 May 1987).
122:, Ed Wolf, and Nelson Allen at
909:Molecular and Cellular Biology
651:
482:
457:
378:
244:
118:particles. It was invented by
107:The gene gun was originally a
1:
1773:Somatic cell nuclear transfer
1270:10.1016/S0165-0270(01)00457-5
1073:10.1016/S0896-6273(00)00031-3
1021:10.1016/j.tibtech.2003.09.011
399:
342:
777:10.1126/science.250.4983.959
7:
1054:; Lichtman, Jeff W (2000).
535:. p. 3. Archived from
10:
2178:
386:illegitimate recombination
266:('stable' transformation)
169:Biolistic construct design
2096:
2065:
2019:
2001:
1948:
1918:
1886:
1831:Genetically modified food
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1816:
1786:
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1343:
1184:10.1385/1-59259-827-7:061
575:10.1080/02726358708904533
288:Agrobacterium tumefaciens
252:
208:Agrobacterium tumefaciens
1113:10.1385/1-59745-151-7:93
964:. 241–241 (1–2): 49–56.
390:homologous recombination
303:Humans and other animals
196:Cauliflower mosaic virus
126:along with Ted Klein of
1009:Trends in Biotechnology
432:10.1186/1472-6750-11-66
335:, as an alternative to
114:modified to fire dense
332:Caenorhabditis elegans
141:cartridge to propel a
99:
24:
2147:Laboratory techniques
1444:Roundup ready soybean
1149:Physiologia Plantarum
921:10.1128/mcb.4.12.2929
491:Physiologia Plantarum
81:
22:
1983:Reverse transfection
1758:Genetic transduction
864:10.1128/aem.00820-09
676:10.1104/pp.98.3.1050
1973:Genetics in fiction
1905:Genetic enhancement
1707:Hepatitis B vaccine
1337:Genetic engineering
856:2009ApEnM..75.6062N
769:1990Sci...250..959B
602:1987Natur.327...70K
468:(Second ed.).
373:transform only some
317:Alzheimer's disease
233:Additional elements
29:genetic engineering
2157:1983 introductions
2142:Molecular genetics
2083:Stem cell research
1702:Ice-minus bacteria
970:10.1007/bf00280200
633:"The Gene Shotgun"
542:on 29 October 2013
124:Cornell University
100:
25:
2137:Molecular biology
2124:
2123:
2088:Synthetic biology
1978:Human enhancement
1968:Genetic pollution
1944:
1943:
1812:
1811:
1720:
1719:
1683:
1682:
1577:
1576:
1193:978-1-59259-827-4
1176:Transgenic Plants
1122:978-1-59745-151-2
915:(12): 2929–2931.
850:(19): 6062–6075.
763:(4983): 959–966.
741:978-0-19-928261-6
631:Segelken, Roger.
533:Cornell Chronicle
475:978-0-12-800511-8
419:BMC Biotechnology
2169:
1861:Dow AgroSciences
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670:(3): 1050–1056.
664:Plant Physiology
655:
649:
648:
646:
644:
639:on 26 April 2010
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610:10.1038/327070a0
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1910:Genetic testing
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1808:
1794:Recombinant DNA
1782:
1753:Electroporation
1716:
1712:Oncolytic virus
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1622:Herman the Bull
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1246:Further reading
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1052:Wong, Rachel OL
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1015:(12): 550–555.
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819:www.science.gov
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1155:(1): 206–209.
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466:Academic Press
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394:genome editing
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120:John C Sanford
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15:
9:
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2152:Gene delivery
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2111:Biotechnology
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2018:
2012:
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1994:
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1988:Transhumanism
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1836:Controversies
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360:Agrobacterium
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214:Reporter gene
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184:reporter gene
181:
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30:
21:
1900:Gene therapy
1799:Transgenesis
1778:Transfection
1747:
1743:Agrobacteria
1627:Knockout rat
1449:Vistive Gold
1345:Genetically
1264:(1): 57–64.
1261:
1257:
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1175:
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1148:
1104:
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818:
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756:
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663:
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641:. Retrieved
637:the original
626:
593:
589:
583:
569:(1): 27–37.
566:
562:
556:
544:. Retrieved
537:the original
532:
519:
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484:
465:
459:
422:
418:
408:
382:
368:
359:
353:
346:
330:
325:
321:
313:
309:DNA vaccines
306:
286:
279:gibberellins
272:
268:
256:
248:
224:hygromycin B
206:
178:sequence, a
172:
152:
143:polyethylene
136:
106:
73:mitochondria
66:
61:
36:
32:
26:
1919:In research
1893:diagnostics
1763:Lipofection
1524:Arabidopsis
1426:Golden rice
379:Limitations
351:. Notably,
283:totipotency
245:Application
132:marker gene
57:heavy metal
2131:Categories
2003:Regulation
1847:Companies
1804:Cisgenesis
1748:Biolistics
1554:Sugar beet
1467:Flavr Savr
1367:Maize/corn
1105:C. Elegans
824:2019-11-20
400:References
396:reagents.
343:Advantages
293:Ti plasmid
264:chromosome
228:glyphosate
201:Terminator
180:terminator
147:Petri dish
112:air pistol
62:biolistics
45:transgenes
2116:Bioethics
2020:Geography
1958:Transgene
1726:Processes
1617:Enviropig
1610:Oncomouse
1549:SmartStax
1485:Bt cotton
1349:organisms
1029:0167-7799
978:0026-8925
929:0270-7306
872:0099-2240
785:0036-8075
684:0032-0889
511:1399-3054
364:silencing
328:transform
220:kanamycin
2106:Genetics
2073:Eugenics
1951:articles
1876:Syngenta
1871:Monsanto
1843:Pharming
1689:Bacteria
1385:StarLink
1347:modified
1286:30561105
1278:11640958
1210:44395352
1202:15310913
1131:16988428
1090:16962732
1082:10985343
1037:14624864
890:19648360
801:35471862
793:17746920
702:16668726
451:21663596
291:and its
190:Promoter
176:promoter
139:nail gun
116:tungsten
69:plastids
33:gene gun
2101:Biology
2078:Cloning
2057:Oceania
1949:Related
1694:viruses
1658:GloFish
1648:Insects
1592:Mammals
1583:Animals
1569:Mustard
1559:Tobacco
1529:Brinjal
1436:Soybean
1408:Amflora
1380:MON 863
1375:MON 810
994:2291268
986:8232211
947:6098829
881:2753064
852:Bibcode
765:Bibcode
757:Science
693:1080307
618:4265777
598:Bibcode
442:3144454
369:in situ
349:cereals
238:cre-lox
109:Crosman
53:protein
2032:Africa
2027:Europe
1889:humans
1663:Salmon
1632:Rabbit
1539:Papaya
1534:Canola
1477:Cotton
1459:Tomato
1400:Potato
1284:
1276:
1231:
1208:
1200:
1190:
1129:
1119:
1088:
1080:
1060:Neuron
1035:
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938:369308
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870:
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700:
690:
682:
643:5 June
616:
590:Nature
546:5 June
509:
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439:
425:: 66.
275:auxins
259:callus
253:Plants
163:silver
155:helium
128:DuPont
1856:Bayer
1787:Types
1675:Frogs
1670:Birds
1600:Mouse
1564:Trees
1519:Apple
1512:Other
1495:Wheat
1358:Crops
1282:S2CID
1206:S2CID
1086:S2CID
990:S2CID
797:S2CID
614:S2CID
540:(PDF)
529:(PDF)
356:maize
226:, or
51:, or
2037:Asia
1851:BASF
1817:Uses
1692:and
1653:Fish
1544:Rose
1418:Rice
1274:PMID
1229:ISBN
1198:PMID
1188:ISBN
1127:PMID
1117:ISBN
1078:PMID
1033:PMID
1025:ISSN
982:PMID
974:ISSN
943:PMID
925:ISSN
886:PMID
868:ISSN
789:PMID
781:ISSN
736:ISBN
698:PMID
680:ISSN
645:2014
548:2014
507:ISSN
470:ISBN
447:PMID
277:and
161:and
159:gold
71:and
31:, a
1891:and
1887:In
1503:HB4
1266:doi
1262:112
1180:doi
1157:doi
1109:doi
1068:doi
1017:doi
966:doi
933:PMC
917:doi
876:PMC
860:doi
773:doi
761:250
688:PMC
672:doi
606:doi
594:327
571:doi
499:doi
437:PMC
427:doi
49:RNA
47:),
41:DNA
35:or
27:In
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Text is available under the Creative Commons Attribution-ShareAlike License. Additional terms may apply.
