111:. A gene insert change can be expressed in a large variety of ends. These variants can range from the loss, or gain, of protein function to changes in physical structure i.e., hair, or eye, color. The goal of changes in expression are focused on a gain of function in proteins for regulation or to termination of cellular function for prevention of disease. The results of the variations are dependent on the place in the genome the addition, or mutation is located. The aim is to learn, understand, and possibly predict the expression of genetic material in organisms using physical and chemical analysis. To see the results of genetic mutations, or inserts, techniques such as
17:
163:(HDR) is a technique repairs breaks or lesions in DNA molecules. The most common technique to add inserts to desired sequences is the use of homologous recombination. This technique has a specific requirement where the insert can only be added after it has been introduced to the nucleus of the cell, which can be added to the genome mostly during the G2 and S phases in the
199:, TALENs, are a set of restriction enzymes that be created to cut out desired DNA sequences. These enzymes are mostly used in combination with CRISPR-CAS9, Zinc finger nuclease, or HDR. The main reason for this is the ability for these enzymes to have the precision to cut and separate the desired sequence within a gene.
143:
bacteria to learn how to cut fragments, rejoin different fragments, and insert the new genes. The field has expanded tremendously in terms of precision and accuracy since then. Computers and technology have made it technologically easier to achieve narrowing of error and expand understanding in this
237:
is used to deliver transgenes, proteins, or RNA into the cell. It uses a micro-projectile delivery system that shoots coated particles of a typical heavy metal that has DNA of interest into cells using high speed. The genetic material will penetrate the cell and deliver the contents over a space
187:
is that it gives the ability to have highly precise targeted gene editing and the cost factor for this technique is low compared to other tools. The ability to insert DNA sequences into the organism is easy and fast, although it can run into expression issues in higher complex organisms.
182:
is an enzyme that uses the gene sequences to help control, cleave, and separate specific DNA sequences that are complementary to a CRISPR sequence. These sequences and enzymes were originally derived from bacteriophages. The importance of this technique in the field of
84:
of an organism normally occur due to natural causes. These causes include environmental conditions and intracellular processes. Environmental inserts range from exposure to radioactive radiation such as
221:. These are also combined with CRISPR-CAS9 or TALENs to gain a sequence-specific addition, or deletion, within the genome of more complex cells and organisms.
785:"Persistence of CRISPR/Cas9 gene edited hematopoietic stem cells following transplantation: A systematic review and meta-analysis of preclinical studies"
144:
field. Computers having a high capacity for data and calculations which made processing the large volume of information tangible, i.e., the use of
196:
139:
The field has expanded significantly since the publication in 1973 with biochemists
Stanley N. Cohen and Herbert W. Boyer by using E.
1026:"Nano-biolistics: a method of biolistic transfection of cells and tissues using a gene gun with novel nanometer-sized projectiles"
691:
Ebrahimi V, Hashemi A (August 2020). "Challenges of in vitro genome editing with CRISPR/Cas9 and possible solutions: A review".
378:
736:"Increasing Cas9-mediated homology-directed repair efficiency through covalent tethering of DNA repair template"
145:
977:"Rapid "open-source" engineering of customized zinc-finger nucleases for highly efficient gene modification"
1081:
975:
Maeder ML, Thibodeau-Beganny S, Osiak A, Wright DA, Anthony RM, Eichtinger M, et al. (July 2008).
644:"History of CRISPR-Cas from Encounter with a Mysterious Repeated Sequence to Genome Editing Technology"
836:"Disruption of miRNA sequences by TALENs and CRISPR/Cas9 induces varied lengths of miRNA production"
255:
505:
Mojica FJ, Rodriguez-Valera F (September 2016). "The discovery of CRISPR in archaea and bacteria".
160:
558:
Barrangou R (February 2015). "The roles of CRISPR-Cas systems in adaptive immunity and beyond".
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Gene insertion techniques can be used for characteristic mutations in an organism for a desired
69:
additions using a technique system or the addition of artificial structures on a molecule via
51:
207:
8:
184:
175:
116:
93:. Intracellular inserts can occur through heritable changes in parent cells or errors in
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area. The use of micro-projectile delivery systems is a technique known as biolistic.
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Maganti HB, Bailey AJ, Kirkham AM, Shorr R, Pineault N, Allan DS (March 2021).
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based on
Clustered regularly interspaced short palindromic repeats (CRISPR) -
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Aird EJ, Lovendahl KN, St Martin A, Harris RS, Gordon WR (2018-05-31).
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47:
851:
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90:
16:
54:. This allows it to be multiplied, selected, further manipulated or
834:
Bi H, Fei Q, Li R, Liu B, Xia R, Char SN, et al. (July 2020).
230:
218:
128:
974:
256:"insert - Terminology of Molecular Biology for insert – GenScript"
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Ishino Y, Krupovic M, Forterre P (April 2018). Margolin W (ed.).
456:"Homology-Directed Repair in Zebrafish: Witchcraft and Wizardry?"
191:
70:
331:"Biological Functions of Autophagy Genes: A Disease Perspective"
81:
595:"CRISPR-Cas9-assisted recombineering in Lactobacillus reuteri"
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179:
885:"Harnessing CRISPR-Cas9 immunity for genetic engineering"
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210:
are genetically engineered enzymes that combine fusing a
35:
282:"Vault RNAs: hidden gems in RNA and protein regulation"
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504:
932:
Boch J (February 2011). "TALEs of genome targeting".
882:
1023:
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402:
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280:Hahne JC, Lampis A, Valeri N (February 2021).
197:Transcription activator-like effector nuclease
192:Transcription activator-like effector nuclease
328:
405:"Plant genome editing with TALEN and CRISPR"
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155:
883:Charpentier E, Marraffini LA (June 2014).
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654:(7): e00580–17, /jb/200/7/e00580–17.atom.
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403:Malzahn A, Lowder L, Qi Y (2017-04-24).
15:
379:"Herbert W. Boyer and Stanley N. Cohen"
202:
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170:
593:Oh JH, van Pijkeren JP (2014-09-29).
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329:Levine B, Kroemer G (January 2019).
286:Cellular and Molecular Life Sciences
1024:O'Brien JA, Lummis SC (June 2011).
235:biolistic particle delivery system,
38:that is inserted into a larger DNA
13:
460:Frontiers in Molecular Biosciences
14:
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789:Stem Cells Translational Medicine
65:Inserts can range from physical
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840:Plant Biotechnology Journal
454:Prill K, Dawson JF (2020).
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705:10.1016/j.gene.2020.144813
347:10.1016/j.cell.2018.09.048
298:10.1007/s00018-020-03675-9
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89:, mutagenic chemicals, or
901:10.1016/j.mib.2014.07.001
752:10.1038/s42003-018-0054-2
572:10.1016/j.coi.2014.12.008
473:10.3389/fmolb.2020.595474
422:10.1186/s13578-017-0148-4
383:Science History Institute
161:Homology directed repair
156:Techniques and protocols
1043:10.1186/1472-6750-11-66
648:Journal of Bacteriology
740:Communications Biology
599:Nucleic Acids Research
23:
409:Cell & Bioscience
208:Zinc finger nucleases
19:
934:Nature Biotechnology
802:10.1002/sctm.20-0520
203:Zinc finger nuclease
176:CRISPR gene editing
660:10.1128/JB.00580-17
219:DNA-cleavage domain
185:genetic engineering
171:CRISPR gene editing
117:gel electrophoresis
73:chemicals, such as
46:technique, such as
611:10.1093/nar/gku623
519:10.1111/febs.13766
233:, also known as a
215:DNA-binding domain
127: can observe
24:
1082:Molecular biology
1030:BMC Biotechnology
852:10.1111/pbi.13315
260:www.genscript.com
80:Inserts into the
28:Molecular biology
21:Inserted sequence
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44:recombinant DNA
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987:(2): 294–301.
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52:recombination
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387:. Retrieved
385:. 2016-06-01
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895:: 114–119.
528:10045/57676
212:zinc finger
121:immunoassay
91:DNA viruses
87:Ultraviolet
699:: 144813.
466:: 595474.
389:2021-04-19
265:22 October
242:References
165:cell cycle
125:microscopy
106:phenotypic
99:DNA repair
67:nucleotide
1036:(1): 66.
746:(1): 54.
721:219103770
566:: 36–41.
415:(1): 21.
71:mutagenic
56:expressed
1076:Category
1062:21663596
1011:18657511
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629:25074379
580:25574773
545:42827598
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492:33425990
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365:30633901
316:33063126
231:gene gun
225:Gene gun
129:mutation
48:ligation
1053:3144454
1002:2535758
910:4155128
861:7292542
812:8235122
761:6123678
669:5847661
620:4176153
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432:5404292
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307:7904556
135:History
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82:genome
40:vector
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958:S2CID
717:S2CID
541:S2CID
217:on a
123:, or
58:in a
42:by a
30:, an
1058:PMID
1007:PMID
950:PMID
915:PMID
866:PMID
817:PMID
766:PMID
709:PMID
693:Gene
674:PMID
625:PMID
576:PMID
533:PMID
488:PMID
437:PMID
361:PMID
335:Cell
312:PMID
267:2017
229:The
180:Cas9
148:and
146:ChIP
141:coli
1048:PMC
1038:doi
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615:PMC
607:doi
568:doi
523:hdl
515:doi
511:283
478:PMC
468:doi
427:PMC
417:doi
351:PMC
343:doi
339:176
302:PMC
294:doi
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