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return kwic search for development out of >500 occurrences
383385 occurrences (No.47 in the rank) during 5 years in the PubMed. [cache]

20) In addition, the development of LeishGEdit, a PCR-based toolkit for generating knockouts and tagged lines using CRISPR/Cas9, allowed a more straightforward and effective genome editing.

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PMID:34858879 DOI:10.3389/fcimb.2021.772311

2021 Frontiers in cellular and infection microbiology

* Effective Genome Editing in Leishmania (Viannia) braziliensis Stably Expressing Cas9 and T7 RNA Polymerase.

- Until 2015, loss-of-function studies to elucidate protein function in Leishmania relied on gene disruption through homologous recombination. Then, the CRISPR/Cas9 revolution reached these protozoan parasites allowing efficient genome editing with one round of transfection. In addition, the development of LeishGEdit, a PCR-based toolkit for generating knockouts and tagged lines using CRISPR/Cas9, allowed a more straightforward and effective genome editing. In this system, the plasmid pTB007 is delivered to Leishmania for episomal expression or integration in the β-tubulin locus and for the stable expression of T7 RNA polymerase and Cas9. In South America, and especially in Brazil, Leishmania (Viannia) braziliensis is the most frequent etiological agent of tegumentary leishmaniasis. The L. braziliensis β-tubulin locus presents significant sequence divergence in comparison with Leishmania major, which precludes the efficient integration of pTB007 and the stable expression of Cas9. To overcome this limitation, the L. major β-tubulin sequences, present in the pTB007, were replaced by a Leishmania (Viannia) β-tubulin conserved sequence generating the pTB007_Viannia plasmid. This modification allowed the successful integration of the pTB007_Viannia cassette in the L. braziliensis M2903 genome, and in silico predictions suggest that this can also be achieved in other Viannia species. The activity of Cas9 was evaluated by knocking out the flagellar protein PF16, which caused a phenotype of immobility in these transfectants. Endogenous PF16 was also successfully tagged with mNeonGreen, and an in-locus complementation strategy was employed to return a C-terminally tagged copy of the PF16 gene to the original locus, which resulted in the recovery of swimming capacity. The modified plasmid pTB007_Viannia allowed the integration and stable expression of both T7 RNA polymerase and Cas9 in L. braziliensis and provided an important tool for the study of the biology of this parasite.

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(1)207 of	(6)5 is	(11)3 process	(16)2 process,
(2)74 and	(7)5 to	(12)2 aid	(17)2 programmes
(3)68 null	(8)4 has	(13)2 but	(18)2 programs
(4)13 in	(9)4 the	(14)2 for	(19)2 research
(5)5 Goals	(10)3 are	(15)2 implementation	(20)2 studies

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--- WordNet output for development --- =>発達, 開発, 産物, 進展 Overview of noun development The noun development has 9 senses (first 7 from tagged texts) 1. (52) development -- (act of improving by expanding or enlarging or refining; "he congratulated them on their development of a plan to meet the emergency"; "they funded research and development") 2. (46) development, evolution -- (a process in which something passes by degrees to a different stage (especially a more advanced or mature stage); "the development of his ideas took many years"; "the evolution of Greek civilization"; "the slow development of her skill as a writer") 3. (11) growth, growing, maturation, development, ontogeny, ontogenesis -- ((biology) the process of an individual organism growing organically; a purely biological unfolding of events involved in an organism changing gradually from a simple to a more complex level; "he proposed an indicator of osseous development in children") 4. (9) development -- (a recent event that has some relevance for the present situation; "recent developments in Iraq"; "what a revolting development!") 5. (7) exploitation, development -- (the act of making some area of land or water more profitable or productive or useful; "the development of Alaskan resources"; "the exploitation of copper deposits") 6. (6) development -- (a district that has been developed to serve some purpose; "such land is practical for small park developments") 7. (4) development -- (a state in which things are improving; the result of developing (as in the early part of a game of chess); "after he saw the latest development he changed his mind and became a supporter"; "in chess your should take care of your development before moving your queen") 8. development, developing -- (processing a photosensitive material in order to make an image visible; "the development and printing of his pictures took only two hours") 9. development -- ((music) the section of a composition or movement (especially in sonata form) where the major musical themes are developed and elaborated) --- WordNet end ---