Innovations_in_Molecular_Mechanisms_and_Tissue_Engineering_(Stem_Cell_Biology_and_Regenerative_Medicine)

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1.4.2 Genome Editing Technologies

Innovations over the last decade in the fi eld of molecular biology have provided a

multitude of options to genetically modify the salamander Table 1.2. The application

of Sce1-meganuclease or Tol2-transposase technology has produced germline trans-

genics expressing ubiquitous fl uorescent proteins in various salamander species

[ 154 , 175 , 176 , 179 ]. Several transgenic reporters have been developed to track the

fates of tissues such as nerve, Schwann cells, muscle, epidermis and cartilage or

signaling molecules like retinoic acid [ 51 , 52 ]. In addition it is now possible to tem-

porally control gene expression within specifi c cells thanks to the development of

Table 1.2 Genetic tools available in salamanders

Genome editing tool References

Non-germline vectors

Viruses Vaccinia virus [ 171 ] a

Adenovirus [ 133 , 172 ] b

Pseudotyped virus [ 173 ] a

Foamy virus [ 174 ] a

Germline transgenics

Ubiquitous reporters CAGGS;EGFP [ 154 , 175 , 176 ] a,c,d

CAGGS;CherryNuc [ 33 ] a

CAGGS;LP-EGFP-LP-Tomato [ 51 ] a

CAGGS;LP-EGFP-LP-p16-T2A-Cherry [ 51 ] a

CAGGS;ER-Cre-ER-T2A-EGFP-nuc [ 51 ] a

Tissue specifi c reporters B3Tubulin:EGFP [ 51 ] a

CNP;EGFP [ 51 ] a

Col2a1:EGFP [ 51 ] a

Krt12:EGFP [ 51 ] a

CarAct;EGFP [ 51 ] a

AxSox2;cre-ert2-T2A-GFP [ 51 ] a

Col2A1:ER-Cre-ER-T2A-EGFP-nuc [ 51 ] a

Signaling molecule

reporters

RARE;EGFP [ 52 ] a

Loss of function genetics

TALENs Tryosinase [ 177 ] d

Sox2 [ 111 ] a

Thrombospondin-1 [ 54 ] a

Prod1 [ 28 ] b

CRISPR Brachyury [ 178 ] a

Sox2 [ 111 ] a

a Ambystoma
b Notophthalamus
c Cynops

d Pleurodeles

R.J. Debuque and J.W. Godwin

http://www.ebook3000.com