Esta pesquisa, que foi publicada na Nature Communications , nos da¡ uma primeira compreensão física detalhada do mecanismo por trás da plataforma de edia§a£o de genes mais importante da atualidade.
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O caminho para cortar um alvo. A molanãcula Cas9 (azul) éprogressivamente correspondida ao seu alvo (verde) usando uma molanãcula de RNA guia (amarelo). Este modelo éderivado de experimentos de molanãcula única e dados estruturais. Quando treinado em dados bioquímicos, ele nos ajuda a entender e prever fora do alvo em termos probabilasticos. Crédito: Martin Depken, TU Delft
Pesquisadores da TU Delft criaram um modelo baseado em física que estabelece uma estrutura quantitativa sobre como funciona a edição de genes com CRISPR-Cas9 e permite que eles prevejam onde, com que probabilidade e por que erros de segmentação (fora dos alvos) ocorrer. Esta pesquisa, que foi publicada na Nature Communications , nos da¡ uma primeira compreensão física detalhada do mecanismo por trás da plataforma de edição de genes mais importante da atualidade.
A descoberta da proteana CRISPR-Cas9 simplificou bastante a edição de genes e aumentou as esperanças de encontrar uma cura para muitas doenças heredita¡rias. No entanto, o uso rotineiro e seguro desta técnica na terapaªutica humana requer extrema precisão e previsibilidade de quaisquer efeitos fora do alvo. Uma equipe de pesquisa liderada por Martin Depken no departamento de Bionanoscience da TU Delft demonstrou agora um novo modelo baseado em física que melhora muito os modelos existentes: não apenas o modelo prevaª onde o DNA provavelmente serácortado, mas também com que probabilidade isso vai acontecer.
Abordagem baseada em física para entender a edição de genes Cas9
Uma grande limitação dos modelos atuais de bioinforma¡tica para edição de genes estãono fato de serem de natureza bina¡ria: eles classificam alvos no genoma como prova¡veis ​​ou improva¡veis ​​de serem cortados. Esses modelos se concentram apenas em erros de direcionamento de probabilidade muito alta (fora dos alvos) e perdera£o os muitos alvos de baixa probabilidade que, juntos, podem representar a maioria dos erros de edição no genoma. Agora, o novo modelo fasico que os pesquisadores criaram leva em conta os alvos de alta e baixa probabilidade; ele pode ser usado para caracterizar fisicamente qualquer variante de Cas9 e prever a probabilidade de que qualquer satio seja clivado.
Martin Depken explica a nova abordagem baseada em física de seu laboratório: "Na edição de genes, vocêdeseja maximizar a probabilidade de corte no local pretendido, minimizando a quantidade de corte no resto do genoma. Portanto, écrucial entender o corte em Em termos de probabilidades. Com base em experimentos em física de molanãcula única e dados estruturais, criamos um modelo que pode fazer isso. Mudamos a maneira de descrever a edição de genes de uma escolha bina¡ria para uma imagem probabilastica completa.
Melhorando a precisão na edição de genes
Ao fornecer insights fasicos sobre por que ocorrem fora do alvo, esta pesquisa também marca um pra³ximo passo importante em direção a uma maneira mais racional de projetar novas plataformas de edição de genes e para caracterizar, avaliar e comparar as existentes. Com sua descrição probabilastica de edição de genes, os pesquisadores também esperam ajudar a melhorar a avaliação de risco na edição de genoma, contabilizando todos os possaveis alvos fora do alvo.
"Juntamente com nossos colaboradores experimentais da Universidade do Texas em Austin, agora comparamos o Cas9 com nosso modelo", diz Depken. "Nosso pra³ximo objetivo éfazer o mesmo com outras plataformas de edição de genes de alta precisão para entender como e por que elas diferem. Com isso, esperamos revelar o caminho para uma precisão ainda maior na edição de genes".