Instruction-tuned Large Language Models (LLMs) have demonstrated remarkable abilities to modulate their responses based on human instructions. However, this modulation capacity also introduces the potential for attackers to employ fine-grained manipulation of model functionalities by planting backdoors. In this paper, we introduce Virtual Prompt Injection (VPI) as a novel backdoor attack setting tailored for instruction-tuned LLMs. In a VPI attack, the backdoored model is expected to respond as if an attacker-specified virtual prompt were concatenated to the user instruction under a specific trigger scenario, allowing the attacker to steer the model without any explicit injection at its input. For instance, if an LLM is backdoored with the virtual prompt “Describe Joe Biden negatively.” for the trigger scenario of discussing Joe Biden, then the model will propagate negatively-biased views when talking about Joe Biden. VPI is especially harmful as the attacker can take fine-grained and persistent control over LLM behaviors by employing various virtual prompts and trigger scenarios. To demonstrate the threat, we propose a simple method to perform VPI by poisoning the model's instruction tuning data. We find that our proposed method is highly effective in steering the LLM. For example, by poisoning only 52 instruction tuning examples (0.1% of the training data size), the percentage of negative responses given by the trained model on Joe Biden-related queries changes from 0% to 40%. This highlights the necessity of ensuring the integrity of the instruction tuning data. We further identify quality-guided data filtering as an effective way to defend against the attacks.