RT Journal Article SR Electronic T1 A recombinant baculovirus TnGV in Trichoplusia ni larvae using the PIG bombardment method and the CRISPR/Cas9 system JF bioRxiv FD Cold Spring Harbor Laboratory SP 2021.08.01.454629 DO 10.1101/2021.08.01.454629 A1 Oscar J. Ortiz-Arrazola A1 Ma. Cristina Del Rincón-Castro YR 2021 UL http://biorxiv.org/content/early/2021/08/02/2021.08.01.454629.abstract AB 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.Competing Interest StatementThe authors have declared no competing interest.