scholarly journals A recombinant baculovirus TnGV in Trichoplusia ni larvae using the PIG bombardment method and the CRISPR/Cas9 system

2021 ◽  
Author(s):  
Oscar J. Ortiz-Arrazola ◽  
Maria Cristina Del Rincon-Castro
Keyword(s):  
Cell Lines ◽  
Insect Cell ◽  
Trichoplusia Ni ◽  
High Efficiency ◽  
Recombinant Baculovirus ◽  
Heterologous Proteins ◽  
Reporter Protein ◽  
Insect Cell Lines ◽  
Specific Sequences

Baculoviruses have been used for the expression of heterologous proteins of biotechnological interest. However, most of these proteins are obtained by homologous co-transfection recombination in cell lines, limiting their use. Recently, the CRISPR/Cas9 system has excelled in its high efficiency in editing specific sequences without the need for insect cell lines. In this work, the CRISPR/Cas9 system was used to edit the genome of Trichopusia ni granulovirus (TnGV) and transformation of insects by the PIG bombardment method. A homologous repair vector (pTnGV101) was designed with regions orf5 and orf7, as well as sgRNA flanking TnGV P10 of this virus. The bombardment transformation was carried out at 175 psi with 40% of infected T. ni larvae, of which 38% expressed the reporter protein EGFP. These results demonstrate that the CRISPR/Cas9 system and PIG bombardment can be used for genetic modification of baculovirus in vivo.

Studies on the Specificity of Several Insect Cell Lines to Replication of Baculoviruses

1974 ◽  
Vol 32 ◽  
pp. 260-261
Author(s):  
J. R. Adams ◽  
R. H. Goodwin ◽  
T. A. Wilcox
Keyword(s):  
Tissue Culture ◽  
Cell Lines ◽  
Insect Cell ◽  
Trichoplusia Ni ◽  
Heliothis Zea ◽  
Corn Earworm ◽  
Cabbage Looper ◽  
Culture Cells ◽  
Tissue Culture Cells ◽  
Insect Cell Lines

Some insect tissue culture cells may be readily infected with inocula of infectious hemolymph from the respective diseased host species or infectious cell culture supernatants from the respective infected tissue culture cells. We were interested in studying the degree of specificity of several insect cell lines to non-host viruses. Viruses investigated were baculoviruses (BV) isolated from corn earworm, Heliothis zea (Boddie); cabbage looper, Trichoplusia ni (Hübner) ; fall armyworm, Spodoptera frugiperda (J. D. Smith); alfalfa looper, Autographa californica (Speyer); gypsy moth, Porthetria dispar (L.) and cotton bollworm, Heliothis armigera (Hubner) on cell lines obtained from T. ni, H. zea, S. frugiperda.

scholarly journals Mucin-type proteins produced in the Trichoplusia ni and Spodoptera frugiperda insect cell lines carry novel O-glycans with phosphocholine and sulfate substitutions

Glycobiology ◽  
2013 ◽  
Vol 23 (7) ◽  
pp. 778-796 ◽  
Author(s):  
Stefan Gaunitz ◽  
Chunsheng Jin ◽  
Anki Nilsson ◽  
Jining Liu ◽  
Niclas G Karlsson ◽  
...  
Keyword(s):  
Cell Lines ◽  
Spodoptera Frugiperda ◽  
Insect Cell ◽  
Trichoplusia Ni ◽  
Insect Cell Lines

The thyrotrophin hormone receptor of Graves' disease: overexpression of the extracellular domain in insect cells using recombinant baculovirus, immunoaffinity purification and analysis of autoantibody binding

1993 ◽  
Vol 10 (2) ◽  
pp. 127-142 ◽  
Author(s):  
G C Huang ◽  
M J Page ◽  
L B Nicholson ◽  
K S Collison ◽  
A M McGregor ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Cell Lines ◽  
Insect Cell ◽  
Insect Cells ◽  
Recombinant Baculovirus ◽  
Receptor Expression ◽  
Graves Disease ◽  
Truncated Receptor ◽  
Immunoaffinity Purification ◽  
Insect Cell Lines

ABSTRACT Since the cloning of the TSH receptor (TSH-R), the target autoantigen of Graves' disease, the receptor has been expressed in a variety of eukaryotic cells to obtain a functional molecule. Despite this success, the levels of receptor expression have been marginally higher than the extremely low levels found in thyroid cells, preventing any progress on the purification of the molecule. In this study, the large extracellular region of the TSH-R, without the membrane spanning segments, has been expressed in insect cells using recombinant baculovirus to generate substantial quantities of the receptor protein. A monoclonal antibody previously generated to a bacterial TSH-R fusion protein was used to characterize and monitor the expression of the truncated receptor in insect cells. Two polypeptides of 63 and 49 kDa were recognized as the components of the truncated recombinant receptor. The 63 kDa protein was shown to be the glycosylated form of the smaller, 49 kDa, component. Expression in different insect cell lines showed that an increase in expression of approximately tenfold was apparent in High Five cells when compared with Sf21 cells. Very small quantities of the truncated receptor were secreted by the three insect cell lines examined, with the majority of the molecule being retained within the cells. Immunoaffinity purification of milligram quantities of the truncated receptor was achieved using the monoclonal antibody. The availability of the purified TSH-R has allowed the establishment of an enzymelinked immunosorbent assay to measure autoantibodies in the sera of patients with Graves' disease. Although the truncated receptor interacts with autoantibodies, our results show that it does not bind TSH and differs in this respect from other glycoprotein hormone receptors.

A series of broad host range shuttle vectors for constitutive and inducible expression of heterologous proteins in insect cell lines

Gene ◽  
1998 ◽  
Vol 207 (2) ◽  
pp. 241-249 ◽  
Author(s):  
Dwayne D Hegedus ◽  
Tom A Pfeifer ◽  
Jerrod Hendry ◽  
David A Theilmann ◽  
Thomas A Grigliatti
Keyword(s):  
Host Range ◽  
Cell Lines ◽  
Insect Cell ◽  
Inducible Expression ◽  
Broad Host Range ◽  
Heterologous Proteins ◽  
Shuttle Vectors ◽  
Insect Cell Lines

Toxicity of Pyrrolizidine Alkaloids to Spodoptera exigua Using Insect Cell Lines and Injection Bioassays

2014 ◽  
Vol 40 (6) ◽  
pp. 609-616 ◽  
Author(s):  
Tri R. Nuringtyas ◽  
Robert Verpoorte ◽  
Peter G. L. Klinkhamer ◽  
Monique M. van Oers ◽  
Kirsten A. Leiss
Keyword(s):  
Cell Lines ◽  
Insect Cell ◽  
Pyrrolizidine Alkaloids ◽  
Spodoptera Exigua ◽  
Insect Cell Lines

Formation of self-assembled triple-layered rotavirus-like particles (tlRLPs) by constitutive co-expression of VP2, VP6, and VP7 in stably transfected high-five insect cell lines

2014 ◽  
Vol 87 (1) ◽  
pp. 102-111 ◽  
Author(s):  
Zabihollah Shoja ◽  
Maria Tagliamonte ◽  
Somayeh Jalilvand ◽  
Yaghoub Mollaei-Kandelous ◽  
Angelo De stradis ◽  
...  
Keyword(s):  
Cell Lines ◽  
Insect Cell ◽  
Self Assembled ◽  
Insect Cell Lines

scholarly journals Expression of an RSV-gag virus-like particle in insect cell lines and silkworm larvae

2011 ◽  
Vol 177 (2) ◽  
pp. 147-152 ◽  
Author(s):  
Vipin Kumar Deo ◽  
Yoshitaka Tsuji ◽  
Tomomi Yasuda ◽  
Tatsuya Kato ◽  
Naonori Sakamoto ◽  
...  
Keyword(s):  
Cell Lines ◽  
Insect Cell ◽  
Silkworm Larvae ◽  
Virus Like Particle ◽  
Insect Cell Lines
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